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Zeitschriftenartikel zum Thema "CAV-3 gene"
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Yu, Wan-cheng, Hai-ying Chen, Hong-li Yang та ін. "rBMSC/Cav-1F92A Mediates Oxidative Stress in PAH Rat by Regulating SelW/14-3-3η and CA1/Kininogen Signal Transduction". Stem Cells International 2019 (28 жовтня 2019): 1–11. http://dx.doi.org/10.1155/2019/6768571.
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Background/Objectives. Carbonic anhydrase 1 (CA1)/kininogen and selenoprotein W (SelW)/14-3-3η signal transduction orchestrate oxidative stress, which can also be regulated by nitric oxide (NO). The mutated caveolin-1 (Cav-1F92A) gene may enhance NO production. This study explored the effect of Cav-1F92A-modified rat bone marrow mesenchymal stem cells (rBMSC/Cav-1F92A) on oxidative stress regulation through CA1/kininogen and SelW/14-3-3η signal transduction in a rat model of monocrotaline- (MCT-) induced pulmonary arterial hypertension (PAH). Method. PAH was induced in rats through the subcutaneous injection of MCT. Next, rBMSC/Vector (negative control), rBMSC/Cav-1, rBMSC/Cav-1F92A, or rBMSC/Cav-1F92A+L-NAME were administered to the rats. Changes in pulmonary hemodynamic and vascular morphometry and oxidative stress levels were evaluated. CA1/kininogen and SelW/14-3-3η signal transduction, endothelial nitric oxide synthase (eNOS) dimerization, and eNOS/NO/sGC/cGMP pathway changes were determined through real-time polymerase chain reaction, Western blot, or immunohistochemical analyses. Results. In MCT-induced PAH rats, rBMSC/Cav-1F92A treatment reduced right ventricular systolic pressure, vascular stenosis, and oxidative stress; downregulated CA1/kininogen signal transduction; upregulated SelW/14-3-3η signal transduction; and reactivated the NO pathway. Conclusions. In a rat model of MCT-induced PAH, rBMSC/Cav-1F92A reduced oxidative stress by regulating CA1/kininogen and SelW/14-3-3η signal transduction.
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Merlini, L. "Familial isolated hyperCKaemia associated with a new mutation in the caveolin-3 (CAV-3) gene." Journal of Neurology, Neurosurgery & Psychiatry 73, no. 1 (2002): 65–67. http://dx.doi.org/10.1136/jnnp.73.1.65.
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Tekin, Hande, and Pınar Edem. "CAV-3-related age-dependent muscle diseases: A novel mutation in mother and son." Neurology Asia 28, no. 3 (2023): 751–55. http://dx.doi.org/10.54029/2023scy.
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The caveolin-3 protein encoded by the CAV-3 gene is a muscle-specific protein found in skeletal, smooth, and cardiac muscle. Caveolin-3 defects lead to several muscle diseases: rippling muscle disease (RMD), limb-girdle muscular dystrophy (LGMD1C), distal myopathy, familial hypertrophic cardiomyopathy, and asymptomatic hyper-CK-emia. While some variants that cause mutations in this gene cause a pure type of disease, some variants may appear as overlap syndromes. Even in the same variants of CAV-3 mutation, the type of muscle disease, its severity, and time of occurrence can be variable. For this reason, it should be known that CAV-3-related diseases and all overlapping diseases can be seen over time, and the patient should be followed up. We report here a 9-year- old boy and his 38-year-old mother who were investigated for asymptomatic hyper-CK-emia and diagnosed with caveolinopathy. The boy had calf hypertrophy and percussion-induced rapid muscle contraction (PIRCs). His mother had calf hypertrophy, contractions due to percussion, and proximal muscle weakness. Mother’s proximal muscles and m. gastrocnemius magnetic resonance imaging (MRI) was normal. The mother had complaints of weakness, showing slow progression starting from the second decade. Heterozygous (ENST000003cav3849.2) c.298A>T p.Ile100Phe variant in exon 2 was detected in the CAV-3 gene. This mutation is classified as pathogenic according to The American College of Medical Genetics and Genomics (ACMG) criteria (PM1, PM2, PP3, PM5). In conclusion, calves’ pseudohypertrophy and mildly raised CK without weakness can be the initial presentation of caveolinopathy. Percussion-induced muscle contractions, rather than muscle rippling, can occur at a young age. The onset of muscle weakness can be delayed during adolescence and can have a slowly deteriorating course associated with myalgia.
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Kalinowski, M., Ł. Adaszek, P. Miłoszowska, et al. "Molecular analysis of a fragment of gene E1B 19K of canine adenovirus 2 (CAV-2) isolated from dogs with symptoms of cough." Polish Journal of Veterinary Sciences 15, no. 3 (2012): 425–30. http://dx.doi.org/10.2478/v10181-012-0066-7.
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AbstractThe aim of this study was to perform molecular analysis of canine adenovirus 2 (CAV-2) E1B 19K gene fragment isolated from 20 dogs of various breeds (12 males and 8 females aged 1-9 years), with clinical symptoms of upper respiratory tract infections, from the Lubelszczyzna region. Nasal swabs were taken from dogs. DNA of CAV-2 was detected using the PCR method in 16 swabs. All PCR products were sequenced, and the obtained sequences were compared with each other and with the sequence of the E1B 19K gene of the CAV-2 strain from an online database of NCBI GenBank: AC 000003. Based on analysis of the obtained sequences, three polymorphic variants of CAV-2 (No.1-3) with homology of 78 - 100% were distinguished. The nucleotide and amino acid sequences of the most frequently represented polymorphic variant, No. 1, differed from the sequences of polymorphic variant No. 2 with one substitution. The nucleotide and amino acid sequence of the E1B 19K gene of CAV-2 AC 000003 differed from the analogous sequences of representatives of variant No. 1 with 44 nucleotide and 19 amino acid substitutions. The small number of nucleotide differences in the E1B 19K CAV-2 gene among the examined own isolates, compared with AC 000003, suggest that the infections in dogs were caused by a relatively genetically stable virus which occurs in eastern Poland.
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Szelechowski, Marion, Annie Fournier, Jennifer Richardson, Marc Eloit, and Bernard Klonjkowski. "Functional organization of the major late transcriptional unit of canine adenovirus type 2." Journal of General Virology 90, no. 5 (2009): 1215–23. http://dx.doi.org/10.1099/vir.0.007773-0.
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Vectors derived from canine adenovirus type 2 (CAV-2) are attractive candidates for gene therapy and live recombinant vaccines. CAV-2 vectors described thus far have been generated by modifying the virus genome, most notably early regions 1 and 3 or the fiber gene. Modification of these genes was underpinned by previous descriptions of their mRNA and protein-coding sequences. Similarly, the construction of new CAV-2 vectors bearing changes in other genomic regions, in particular many of those expressed late in the viral cycle, will require prior characterization of the corresponding transcriptional units. In this study, we provide a detailed description of the late transcriptional organization of the CAV-2 genome. We examined the major late transcription unit (MLTU) and determined its six families of mRNAs controlled by the putative major late promoter (MLP). All mRNAs expressed from the MLTU had a common non-coding tripartite leader (224 nt) at their 5′ end. In transient transfection assays, the predicted MLP sequence was able to direct luciferase gene expression and the TPL sequence yielded a higher amount of transgene product. Identification of viral transcriptional products following in vitro infection confirmed most of the predicted protein-coding regions that were deduced from computer analysis of the CAV-2 genome. These findings contribute to a better understanding of gene expression in CAV-2 and lay the foundation required for genetic modifications aimed at vector optimization.
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He, Miaomiao, Jie Qiu, Yan Wang, Yang Bai, and Guangzhi Chen. "Caveolin-3 and Arrhythmias: Insights into the Molecular Mechanisms." Journal of Clinical Medicine 11, no. 6 (2022): 1595. http://dx.doi.org/10.3390/jcm11061595.
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Caveolin-3 is a muscle-specific protein on the membrane of myocytes correlated with a variety of cardiovascular diseases. It is now clear that the caveolin-3 plays a critical role in the cardiovascular system and a significant role in cardiac protective signaling. Mutations in the gene encoding caveolin-3 cause a broad spectrum of clinical phenotypes, ranging from persistent elevations in the serum levels of creatine kinase in asymptomatic humans to cardiomyopathy. The influence of Caveolin-3(CAV-3) mutations on current density parallels the effect on channel trafficking. For example, mutations in the CAV-3 gene promote ventricular arrhythmogenesis in long QT syndrome 9 by a combined decrease in the loss of the inward rectifier current (IK1) and gain of the late sodium current (INa-L). The functional significance of the caveolin-3 has proved that caveolin-3 overexpression or knockdown contributes to the occurrence and development of arrhythmias. Caveolin-3 overexpression could lead to reduced diastolic spontaneous Ca2+ waves, thus leading to the abnormal L-Type calcium channel current-induced ventricular arrhythmias. Moreover, CAV-3 knockdown resulted in a shift to more negative values in the hyperpolarization-activated cyclic nucleotide channel 4 current (IHCN4) activation curve and a significant decrease in IHCN4 whole-cell current density. Recent evidence indicates that caveolin-3 plays a significant role in adipose tissue and is related to obesity development. The role of caveolin-3 in glucose homeostasis has attracted increasing attention. This review highlights the underlining mechanisms of caveolin-3 in arrhythmia. Progress in this field may contribute to novel therapeutic approaches for patients prone to developing arrhythmia.
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Pallister, Jackie, Kevin J. Fahey, and Michael Sheppard. "Cloning and sequencing of the chicken anaemia virus (CAV) ORF-3 gene, and the development of an ELISA for the detection of serum antibody to CAV." Veterinary Microbiology 39, no. 1-2 (1994): 167–78. http://dx.doi.org/10.1016/0378-1135(94)90097-3.
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Robinson, J. M. "What Can Epitope Specific Antibodies Tell us About the Organization of Caveolin in Cells?" Microscopy and Microanalysis 7, S2 (2001): 1030–31. http://dx.doi.org/10.1017/s1431927600031226.
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There are three members of the caveolin (CAV) gene family that give rise to four polypeptides. These polypeptides are CAV-1α, CAV-1β, CAV-2, and CAV-3. The CAV-1β isoform is a truncated form of CAV-1α that lacks 31 amino acids at the N-terminus of the molecule. The CAV- 1β molecule arises through an alternative splicing mechanism.Caveolae are specialized plasma membrane microdomains that are expressed at high levels in some cell types (e.g., endothelium, adipocytes, fibroblasts). These specialized regions of the plasma membrane have a characteristic omega-shaped appearance with diameters ranging from 40-90 run. They are distinct from clathrin-coated pits since they lack the characteristic coated appearance in electron microscopy. Caveolae were among the first structures to be discovered by biological electron microscopy. However, biochemical characterization of these structures did not begin in earnest until a marker protein was identified. The initial marker was the 22-kDa protein known as caveolin.
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Smuts, Heidi E. M. "Novel Gyroviruses, including Chicken Anaemia Virus, in Clinical and Chicken Samples from South Africa." Advances in Virology 2014 (2014): 1–7. http://dx.doi.org/10.1155/2014/321284.
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Introduction. Chicken anaemia virus, CAV, was until recently the only member of theGyrovirusgenus. 6 novel gyroviruses, AGV2, HGyV1, and GyV3-6, have since been discovered in human and chicken samples.Methods. PCR amplification of the VP2 gene was used to detect AGV2/HGyV1, GyV3, and CAV in a range of clinical samples including stool, respiratory, CSF, and HIV-positive plasma. Screening of fresh local chicken meat was also performed.Results. AGV2/HGyV1 or GyV3 was detected in stools from healthy children (17/49, 34.7%) and patients with diarrhoea (22/149, 14.8%). 1.2% (3/246) nasopharyngeal respiratory samples were positive. No AGV2/HGyV1 or GyV3 was detected in nasal swabs from wheezing patients, in CSF from patients with meningitis, and in HIVpositive plasma. CAV was found in 51% (25/49) of stools from healthy children and 16% (24/149) in diarrhoea samples. Screening of 28 chicken samples showed a higher prevalence of gyrovirus (20/28, 71%) compared to CAV (1/28, 3.6%). Phylogenetic analysis of the CAV VP1 gene showed South African sequences clustering with Brazilian isolates from genotypes D2 and A2.Conclusion. Novel gyroviruses, including CAV, are present in the South African population with diarrhoea and respiratory illness as well as in healthy children. Their presence suggests an origin from chicken meat consumption.
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Nogueira, E. O., L. Brentano, and A. J. P. Ferreira. "A VP3/VP1 gene polymerase chain reaction assay for detection of chicken anemia virus in broiler samples." Arquivo Brasileiro de Medicina Veterinária e Zootecnia 57, suppl 2 (2005): 131–40. http://dx.doi.org/10.1590/s0102-09352005000800001.
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A PCR assay was designed for amplification of the highly conserved VP3 gene and a 5' region of the VP1 gene, for the diagnosis of CAV in organ samples of broiler flocks suspected of chicken infectious anemia. A comparison of the VP3/VP1 PCR with in vivo virus isolation revealed 100% agreement of the results, with 13 positive and 3 negative samples in both assays, indicating that the VP3/VP1 PCR is a specific diagnostic method. Tissues from additional 24 broiler chicken flocks, with CAV-like lesions and clinical history were then tested only by the VP3/VP1 PCR and a reference PCR with published primers for the VP1 gene. Nineteen samples resulted positive and one negative in both PCR, while another 4 samples were positive only in the VP3/VP1 PCR. These results indicate that the VP3/VP1 PCR is a sensitive, specific diagnostic test, suitable as an alternative to the expensive and time consuming in vivo virus isolation method, specially considering the difficult diagnosis of CAV strains not readily adaptable to MSB-1 cell culture.
Dissertationen zum Thema "CAV-3 gene"
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Lemerle, Eline. "Rôle des cavéoles dans la formation des tubules-T et dans la physiopathologie des cavéolinopathies." Electronic Thesis or Diss., Sorbonne université, 2021. https://accesdistant.sorbonne-universite.fr/login?url=https://theses-intra.sorbonne-universite.fr/2021SORUS010.pdf.
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Dans le muscle squelettique, des invaginations du sarcolemme appelées cavéoles et leur composant principal Cav-3 seraient impliqués dans la formation des tubules transverses, des structures musculaires permettant de propager le potentiel d'action dans la fibre musculaire. Pourtant, ce mécanisme demeure à ce jour inconnu. L’importance des cavéoles et de Cav-3 est accentuée par l’existence de défauts dans l’organisation et la fonction des cavéoles dans le cas de cavéolinopathies, des maladies neuromusculaires autosomiques dominantes dues à des mutations dans le gène CAV-3 et dont les mécanismes physiopathologiques sont à ce jour incompris. L’objectif de mon projet était de comprendre le rôle des cavéoles dans la formation précoce des tubules-T. Une technique de microscopie corrélative combinant de la fluorescence à super résolution et de la microscopie électronique sur répliques de métal a permis d’examiner en détails les composants moléculaires des cavéoles et des tubules-T dans des myotubes extensivement différenciés. J’ai ainsi montré l'organisation des cavéoles sur des plateformes de Bin1 formant ainsi une nouvelle structure en anneaux semblant optimiser la tubulation de la membrane afin d’initier la formation des tubules-T. Ces anneaux ainsi que la tubulation des membranes par Bin1 sont altérés dans le cas de défauts d’expression de Cav-3 et dans les myotubes de patients cavéolinopathes. Mes travaux suggèrent que les anneaux de cavéoles constituent le site d’initiation des tubules-T et apportent les bases d’une caractérisation de la biogénèse des tubules-T dans le muscle squelettique et dans la physiopathologie des cavéolinopathies<br>In skeletal muscle, invaginations of the sarcolemma called caveolae and their main component Cav-3 are thought to be involved in the formation of transverse tubules. T-tubules are muscle structures that allow the action potential to be propagated into the muscle fibre. However, the mechanism linking them together remains unknown. The importance of caveolae and Cav-3 is accentuated by the existence of defects in the organisation and function of caveolae in caveolinopathies, autosomal dominant neuromuscular diseases due to mutations in the CAV-3 gene, the pathophysiological mechanisms of which are still not understood. The objective of my project was to understand the role of caveolae in the early formation of T-tubules. A correlative microscopy technique combining super-resolution fluorescence and electron microscopy on metal replicas was used to examine in detail the molecular components of caveolae and T-tubules in extensively differentiated myotubes. I showed the organisation of caveolae on Bin1 platforms forming a novel ring structure that appears to optimise membrane tubulation in the initiation of T-tubule formation. These rings and Bin1-mediated membrane tubulation are impaired in Cav-3 expression defects and in myotubes from caveolinopathic patients. My work suggests that caveolae rings are the site of T-tubule initiation and provides the basis for characterising T-tubule biogenesis in skeletal muscle and in the pathophysiology of caveolinopathies
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Song, Xiaomin. "Distribution and molecular characterization of 3'-UTR binding proteins specific to the (U)¦1¦5 region of Cas-1 gene." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1997. http://www.collectionscanada.ca/obj/s4/f2/dsk2/tape16/PQDD_0013/MQ28664.pdf.
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Stanojevic, Nadia. "The yeast cap-binding complex is co-transcriptionally recruited to the ADH1 gene and is not required for recruitment of the elongation, RNA 3'-end formation and export machineries in vivo /." Available to subscribers only, 2005. http://proquest.umi.com/pqdweb?did=1079658481&sid=3&Fmt=2&clientId=1509&RQT=309&VName=PQD.
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Kiss, Andor Joseph. "A spatial and temporal study of mRNA binding proteins specific to a 134 base CA dinucleotide-rich portion of the 3'-untranslated region from the BALB/c mouse catalase gene, Cas-1." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1997. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp04/mq28596.pdf.
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Novak, Gabriela. "Upregulation of CaMKIIβ and Nogo-C mRNA in Schizophrenia and the Prevalence of CAA Insert in the 3’UTR of the Nogo Gene". Thesis, 2008. http://hdl.handle.net/1807/11240.
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Schizophrenia may result from altered gene expression leading to abnormal neurodevelopment. In a search for genes with altered expression in schizophrenia, cDNA library subtractive hybridization experiments using post-mortem human frontal cerebral cortices from schizophrenia individuals and neurological controls were performed.
I found the mRNA of two neurodevelopmentally important genes, Nogo (RTN4) and calcium/calmodulin-dependent protein kinase II beta (CaMKIIβ), to be overexpressed in post-mortem frontal cortex tissues from patients who suffered with schizophrenia. I used the quantitative real-time polymerase chain reaction method to determined the mRNA levels of these genes in tissues from age- and sex-matched individuals.
Nogo is a myelin-associated protein which inhibits the outgrowth of neurites and nerve terminals. The gene produces three splice variants, Nogo-A, B and C. I found Nogo-C mRNA to be overexpressed by 26% in schizophrenia. I also found a 17% reduction of Nogo-B mRNA in samples from individuals who had been diagnosed with severe depression. Furthermore, I showed that there is a direct correlation between the expression of both Nogo-A and -C and the presence of a CAA insert in the 3’UTR of the Nogo gene.
CaMKII is a kinase localized at the postsynaptic density. The holoenzyme is primarily composed of the subunits α and β, encoded by two separate genes. It influences the expression of many neuroreceptors, in particular receptors of the glutamatergic pathway. CaMKII also mediates neural maturation during puberty, a time of onset of schizophrenia. The expression of CaMKIIα was elevated 29% in frontal cortex tissues of patients who suffered from depression. The expression of CaMKIIβ was elevated 27% in tissues of schizophrenia patients and 36% in tissues of patients diagnosed with depression. Upregulation of CaMKIIβ was associated with the presence of the CAA insert in at least one copy of the Nogo gene in a group containing both healthy subjects and patients with mental illness, possibly linking the CaMKII and Nogo pathways. The values for the expression of Nogo, CaMKIIα and CaMKIIβ were normalized to β-glucuronidase expression to minimize the effects of mRNA degradation. These results confirm that upregulation of Nogo-C and CaMKIIβ is likely associated with schizophrenia.
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Rodriguez, Angel E. "Characterization of CAL 27 and HSC-3 cell lines. DPAGT1 gene expression and association with oral squamous cell carcinoma genesis and metastasis." Thesis, 2016. https://hdl.handle.net/2144/18669.
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Cancer, a disease of an uncontrolled cell division, growth and metastasis as a result of genetic mutations, environmental factors and host response, is affecting populations worldwide. Etiology, pathogenicity, and genetics related to cancer are not well understood, and treatment has not been as effective as scientists have expected. Continual research is being done to improve current understanding and treatments.
Oral squamous cell carcinoma (OSCC) is one of the most common head and neck cancers (representing >90 % of all head and neck cancers) involving neoplasms of the oral cavity and oropharynx. OSCC is a very pernicious malignancy developed from epithelial cells. There is evidence that a key N-glycosylation gene, DPAGT1, is associated with cancer. Although N-glycosylation of proteins is involved in organ development and homeostasis of tissue, overexpression of DPAGT1 has been implicated in oral cancer initiation and metastasis. Defects in N-glycosylation underlie congenital disorders, while hyper-N-glycosylation has been shown to be a feature of many cancers.
The N-glycosylation pathway directs cell adhesion and cytoskeletal dynamics by impacting the function of E-cadherin, a major epithelial cell-cell adhesion receptor. E-cadherin is a tumor suppressor responsible for the organization of multiprotein complexes named adherens junctions (AJs). In epithelial cells, stable AJs are essential for several cellular processes, including inhibition of cell proliferation, reorganization of the actin cytoskeleton, and maintenance of an epithelial phenotype. Indeed, restoration of AJs has been shown to revert cancer cells from a mesenchymal to an epithelial phenotype and to reduce invasiveness. Previous work has shown that upregulation of DPAGT1 plays a pivotal role in driving canonical WNT/β-catenin signaling (also known as canonical Wnt signaling) that represses E-cadherin adhesions and drives tumorigenic phenotypes in oral cancer. This suggests a role in coordinating balance between proliferation and adhesion by DPAGT1.
To date, little is known about the molecular and cellular details underlying differences among OSCC cell lines. CAL 27 and HSC-3 are human cancer cell lines commonly used to in laboratory OSCC research. The main differences between these cell lines include capsular tumors formed by CAL27 cells in nude mouse models in contrast to non-capsular and invasive tumors formed by HSC-3 cells. The goal of this study was to characterize biochemical differences between these two cell lines for further research.
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Kellner, Tobias [Verfasser]. "Expressionsanalyse der potenziellen Tumor-Suppressor-Gene H-REV107-2/TIG 3 & MUC18/Mel-CAM in humanen Ovarialkarzinom-Zelllinien und Ovarialkarzinomen / von Tobias Kellner." 2007. http://d-nb.info/988265400/34.
Der volle Inhalt der QuelleBücher zum Thema "CAV-3 gene"
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Joyner, Alexandra, ed. Gene Targeting. Oxford University Press, 1999. http://dx.doi.org/10.1093/oso/9780199637928.001.0001.
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Since the publication of the first edition of Gene Targeting: A Practical Approach in 1993 there have been many advances in gene targeting and this new edition has been thoroughly updated and rewritten to include all the major new techniques. It provides not only tried-and-tested practical protocols but detailed guidance on their use and applications. As with the previous edition Gene Targeting: A Practical Approach 2e concentrates on gene targeting in mouse ES cells, but the techniques described can be easily adapted to applications in tissue culture including those for human cells. The first chapter covers the design of gene targeting vectors for mammalian cells and describes how to distinguish random integrations from homologous recombination. It is followed by a chapter on extending conventional gene targeting manipulations by using site-specific recombination using the Cre-loxP and Flp-FRT systems to produce 'clean' germline mutations and conditionally (in)activating genes. Chapter 3 describes methods for introducing DNA into ES cells for homologous recombination, selection and screening procedures for identifying and recovering targeted cell clones, and a simple method for establishing new ES cell lines. Chapter 4 discusses the pros and cons or aggregation versus blastocyst injection to create chimeras, focusing on the technical aspects of generating aggregation chimeras and then describes some of the uses of chimeras. The next topic covered is gene trap strategies; the structure, components, design, and modification of GT vectors, the various types of GT screens, and the molecular analysis of GT integrations. The final chapter explains the use of classical genetics in gene targeting and phenotype interpretation to create mutations and elucidate gene functions. Gene Targeting: A Practical Approach 2e will therefore be of great value to all researchers studying gene function.
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Wade, Tracey D., and Cynthia Bulik. Genetic Influences on Eating Disorders. Edited by W. Stewart Agras and Athena Robinson. Oxford University Press, 2017. http://dx.doi.org/10.1093/oxfordhb/9780190620998.013.5.
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The current chapter reviews our progress in understanding how genes influence eating disorders by addressing the following areas: (1) how recognition of genetic influences on eating disorders emerged; (2) the complexities of gene environment interplay; (3) what twin studies can tell us about gene environment interplay, and (4) the current state of molecular genetic studies. It is concluded that both genes and nonshared environment play a critical role in the explanatory framework for the etiology of eating disorders. Shared environment is likely to contribute to the development of cognition and attitudes that may initiate disordered eating practices. Researchers are on the cusp of identifying specific genes that are implicated, and explication of the manner in which genes and the environment work together to increase risk for eating disorders hinges on the collection of larger samples.
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Dode, L. Identification and Characterisation of the Human Sarco/ Endoplasmic Reticulum Ca2+-Atpase 3 (Serca3) Gene. Leuven University Press, 1998.
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Schrag, Brian, and Kathleen J. Van Buren. Connect Goals to Genres. Oxford University Press, 2018. http://dx.doi.org/10.1093/oso/9780190878276.003.0004.
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Step 3 follows Step 2, in which community members choose a goal for artistic programs. Step 3 provides guidance for choosing the desired effects of arts programs; choosing the content of arts programs; choosing a genre or genres that can communicate the content and produce the desired effects; and imagining events that could include the performance of new artistic works. Specific suggestions and tables (such as the Genre Comparison Chart) assist readers in moving through these processes. This step also considers how communication occurs through an artistic event. Step 3 concludes with a note on tourism and other repurposing of arts.
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Maj, Dorota. Modyfikujący wpływ roślinnych dodatków paszowych na użytkowość mięsną i ekspresję wybranych genów u królików w zależności od wieku i płci. Publishing House of the University of Agriculture in Krakow, 2017. http://dx.doi.org/10.15576/978-83-66602-29-8.
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The aim of the study was to determine the effect of feed additives (algae, soybean, and sunflower oil) used in the rabbit feed on: growth indices and slaughter traits, pH, colour, texture, chemical composition, fatty acid profile and oxidative stability (TBARS) of the meat as well as FTO and FABP4 genes expression in the meat’s intramuscular fat (m. longissimus lumborum), depending on the age and sex. The experimental material consisted of Termond White rabbits (n = 160, 80 females and 80 males). Animals were weaned on the 35th day of life, and housed in metal cages arranged in batteries (4 rabbits of the same sex in a cage). From weaning to 12 or 18 weeks of age, the rabbits were fed pellets ad libitum. Animals in the control group (C) received non-supplemented pellets throughout the experiment. In the other groups, the pellet contained 1% algae (A), 3% sunflower oil (OS), and 3% soybean oil(SO).The experimental diets were formulated to have similar protein and energy content. Diets were balanced by lowering the proportion of other feed components. The total share of all components remained at 100%. The results indicate that 3% vegetable oils (soybean or sunflower) supplementation of diets for growing rabbits leads to an increase of body weight and improvement of some of the slaughter traits, while 1% addition of algae to the feed causes deterioration of body weight and slaughter traits. The effect of oil additive depends on the animals’ age. Supplementation of the rabbits’ diet with algae (1%) or sunflower and soybean oils (3%) led to an increase in the dressing percentage of rabbits slaughtered at 18 weeks of age (approx. 3%), but had no effect on the dressing percentage of rabbits slaughtered at 12 weeks of age. Feeding pellets with either 3% vegetable oils or 1% algae additive to the rabbits did not significantly change the chemical composition of the meat. Protein content increased and intramuscular fat content decreased with age, while ash and water content were similar. The feed additives significantly differentiated meat acidity without deteriorating meat quality. Diet modification has not affected negatively meat colour. 24 h after the slaughter, the colour of rabbit meat was similar across the studied feeding groups. Correlation between diet and rabbits’ age was found. Meat texture (hardness, springiness and chewiness) of all rabbit groups slaughtered at 12 weeks of age was similar, and the shear for cewas greater in rabbits fed pellets with algae and soybean oil. At 18 weeks of age, rabbit meat from experimental groups had lower hardness and chewiness, compared to meat of the animals from the control group. Meat shear force was higher in the control group, and from algae-supplemented group. The correlation between diet and age was also found. The use of 3% vegetable oils or 1% algae as feed additives significantly reduced meat oxidative stability. Soybean or sunflower oil (3%) usedas feed additives favourably modified the fatty acid composition of intramuscular fat. Polyunsaturated fatty acids (PUFA) content was increased, including linoleic acid, and PUFA/MUFA ratio was improved. The content of these acids decreased with age. The use of algae (1%) as a feed additive resulted in positive effect on the increase of n-3 fatty acid content (EPA and DHA) in meat intramuscular fat. Algae supplementation improved pro-health properties of meat, with low n-6/n-3 acid ratio (2.5), indicating that diet modification may affect the fatty acid composition of rabbit meat. The influence of diet and age on FTO and FABP4 gene expression in meat intramuscular fat (m. longissimus lumborum) was found. FTO and FABP4 gene expression increased with age and was the highest in the group of rabbits with 1% algae supplementation in the diet. The effect of rabbits’ gender on growth, slaughter traits, meat quality and gene expression in rabbits was not observed. In conclusion, the use of natural feed additives, such as sunflower, soybean oil or algae, can improve the nutritional value of rabbit meat, without changing its chemical or physical properties, and therefore the meat can serve as functional food, with properties beneficial to human health. The results obtained in this study also indicate that the expression of FTO and FABP4 genes in rabbit muscles is regulated by dietary factors and age, which, in addition to cognitive significance, has practical implications for improving technological and dietary quality of rabbit meat.
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Kirchman, David L. Predation and protists. Oxford University Press, 2018. http://dx.doi.org/10.1093/oso/9780198789406.003.0009.
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Protists are involved in many ecological roles in natural environments, including primary production, herbivory and carnivory, and parasitism. Microbial ecologists have been interested in these single-cell eukaryotes since Antonie van Leeuwenhoek saw them in his stool and scum from his teeth. This chapter focuses on the role of protozoa (purely heterotrophic protists) and other protists in grazing on other microbes. Heterotrophic nanoflagellates, 3–5 microns long, are the most important grazers of bacteria and small phytoplankton in aquatic environments. In soils, flagellates are also important, followed by naked amoebae, testate amoebae, and ciliates. Many of these protists feed on their prey by phagocytosis, in which the prey particle is engulfed into a food vacuole into which digestive enzymes are released. This mechanism of grazing explains many factors affecting grazing rates, such as prey numbers, size, and composition. Ingestion rates increase with prey numbers before reaching a maximum, similar to the Michaelis–Menten equation describing uptake as a function of substrate concentration. Protists generally eat prey that are about ten-fold smaller than they are. In addition to flagellates, ciliates and dinoflagellates are often important predators in the microbial world and are critical links between microbial food chains and larger organisms Many protists are capable of photosynthesis. In some cases, the predator benefits from photosynthesis carried out by engulfed, but undigested photosynthetic prey or its chloroplasts. Although much can be learnt from the morphology of large protists, small protists (<10 μm) often cannot be distinguished by morphology, and as seen several times in this book, many of the most abundant and presumably important protists are difficult to cultivate, necessitating the use of cultivation-independent methods analogous to those developed for prokaryotes. Instead of the 16S rRNA gene used for bacteria and archaea, the 18S rRNA gene is key for protists. Studies of this gene have uncovered high diversity in natural protist communities and, along with sequences of other genes, have upended models of eukaryote evolution. These studies indicate that the eukaryotic Tree of Life consists almost entirely of protists, with higher plants, fungi, and animals as mere branches.
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Laland, Kevin N., and Gillian R. Brown. The Social Construction of Human Nature. Oxford University Press, 2018. http://dx.doi.org/10.1093/oso/9780198823650.003.0008.
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What is the job that the term ‘human nature’ is expected to do? Three notions are prevalent but are problematic: (1) Distinguishing what is biological from what is cultural/environmental. Here the term fails. (2) Characterizing the defining features of humanity, thereby allowing us to be distinguished from other species. This stance is tenable but contributes little. (3) Characterizing what is universal or typical about humanity, because of our ‘evolved biological heritage’. Here the term is tenable but misleading and hence counterproductive. ‘Human nature’ is equally reciprocally caused by gene–culture coevolution and niche construction. Given that the term has little explanatory power but carries extensive baggage, we suggest that it should be abandoned. It can be replaced with descriptions of human behaviour and cognition as the product of socially mediated internal and external constructive processes operating over both developmental and evolutionary timescales.
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Muir, Stephanie. Studying City of God. Liverpool University Press, 2008. http://dx.doi.org/10.3828/liverpool/9781903663592.001.0001.
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A leading example of a resurgent Latin American cinema — ‘la buena onda’ — in the early twenty-first century, City of God was a huge international popular and critical success. A combination of intoxicating, Hollywood-style genre film-making and hard-hitting, social-realist subject matter, it was hailed as a masterpiece at Cannes in 2002 and seen by over 3 million people in Brazil, including the Brazilian cabinet. This book considers: The historical and industrial context of City of God — a brief history of Latin American cinema is followed by a more detailed account of film-making in Brazil — from light-hearted travelogues to Cinema Novo and after — all in the context of increasing globalisation; Narrative and Genre — how the film uses the components of narrative in a complex way, experimentally manipulating time while using traditional genre conventions that are highly recognisable to mainstream audiences; Film language — the formal elements of the film are dissected through a detailed illustrated analysis of the kinetic, scene setting opening sequence; Audience responses — from establishment critical reaction to fan-based internet sites and student feedback; Representation and Ideology — just how ‘authentic’ can a film such as City of God hope to be? Does its style overwhelm its subject matter?
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Harding, Sian E. The Exquisite Machine. The MIT Press, 2022. http://dx.doi.org/10.7551/mitpress/12836.001.0001.
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How science is opening up the mysteries of the heart, revealing the poetry in motion within the machine. Your heart is a miracle in motion, a marvel of construction unsurpassed by any human-made creation. It beats 100,000 times every day—if you were to live to 100, that would be more than 3 billion beats across your lifespan. Despite decades of effort in labs all over the world, we have not yet been able to replicate the heart's perfect engineering. But, as Sian Harding shows us in The Exquisite Machine, new scientific developments are opening up the mysteries of the heart. And this explosion of new science—ultrafast imaging, gene editing, stem cells, artificial intelligence, and advanced sub-light microscopy—has crucial, real-world consequences for health and well-being. Harding—a world leader in cardiac research—explores the relation between the emotions and heart function, reporting that the heart not only responds to our emotions, but it also creates them. The condition known as Broken Heart Syndrome, for example, is a real disorder that can follow bereavement or stress. The Exquisite Machine describes the evolutionary forces that have shaped the heart's response to damage, the astonishing rejuvenating power of stem cells, how we can avoid heart disease, and why it can be so hard to repair a damaged heart. It tells the stories of patients who have had the devastating experiences of a heart attack, chaotic heart rhythms, or stress-induced acute heart failure. And it describes how cutting-edge technologies are enabling experiments and clinical trials that will lead us to new solutions to the worldwide scourge of heart disease.
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Branham, R. Bracht. Inventing the Novel. Oxford University Press, 2019. http://dx.doi.org/10.1093/oso/9780198841265.001.0001.
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Bakhtin as a philosopher and a student of the novel is intent upon the novel’s role in the history of consciousness. His project fails if he is wrong about the dialogic nature of consciousness or the cultural centrality of the novel as the only discourse that can model human consciousness and its intersubjective character. Inventing the Novel is an argument in four stages: the Introduction surveys Bakhtin’s life and his theoretical work in the 1920s, which grounded his work on the novel, as investigated in following chapters. Chapter 1 sketches Bakhtin’s view of literary history as an agonistic dialogue of genres, concluding with his claim that the novel originates as a new way of evaluating time. Chapter 2 explores Bakhtin’s theory of chronotopes: how do forms of time and space in ancient fiction delimit the possible representation of the human? Chapter 3 assesses Bakhtin’s poetics of genre in his account of Menippean satire as crucial in the history of the novel. Chapter 4 uses Petronius to address the prosaics of the novel, exploring Bakhtin’s account of how novelists of “the second stylistic line” orchestrate the babble of voices expressive of an era into “a microcosm of heteroglossia,” focusing it through the consciousness of characters “on the boundary” between I and thou. Insofar as this analysis succeeds, it evinces the truth of Bakhtin’s claim that the role of Petronius’s Satyrica in the history of the novel is “immense.”
Buchteile zum Thema "CAV-3 gene"
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Ross, Samuel E., and Ozren Bogdanovic. "Generation and Molecular Characterization of Transient tet1/2/3 Knockouts." In Methods in Molecular Biology. Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1294-1_17.
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Abstract5-methylcytosine (5mC) is a gene-regulatory mark associated with transcriptional repression. 5mC can be erased through the catalytic action of Ten-eleven translocation (TET) methylcytosine dioxygenases (TET1, TET2, TET3), which oxidize 5mC resulting in its removal from the genome. In vertebrates, TET enzymes facilitate DNA demethylation of regulatory regions linked to genes involved in developmental processes. Consequently, TET ablation leads to severe morphological defects and developmental arrest. Here we describe a system that can facilitate the study of relationships between TET enzymes, 5mC, and embryo development. We provide detailed descriptions for the generation of F0 zebrafish tet1/2/3 knockouts using CRISPR/Cas9 technology and elaborate on the strategies to assess the impact of TET loss by reduced representation bisulfite sequencing (RRBS).
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Zhang, Lei, and Jian-Jun Hu. "Transgenic poplar gene flow monitoring in China." In Gene flow: monitoring, modeling and mitigation. CABI, 2021. http://dx.doi.org/10.1079/9781789247480.0004.
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Abstract Poplar is cultivated widely for pulpwood, firewood, and timber. Transgenic poplar may be part of a solution for wood demand in China. Because transgene escape is an important part of ecological security evaluation of transgenic plants, in this chapter we discuss a real transgenic poplar case study. In this case study, mature transgenic male Populus nigra plants harbored a Bacillus thuringiensis toxin gene (i.e. Bt poplar). A plantation of these plants served as a testbed for a relevant example for gene flow monitoring in China. Furthermore, we discuss environmental risk assessment (ERA) of these transgenic plants. While transgenes can drift to related species through natural and controlled pollination, the probability of transgene drift appears to be very low in the field. The resultantBt poplar seeds occurred at a frequency from about 0.15% at 0 m to about 0.02% at 500 m away from the Bt poplar. The Bt poplar progeny seeds had decreased germination within 3 weeks in the field (from 68% to 0%), compared with the 48% germination rate after 3 weeks at 4°C. The survival rate of seedlings in the field was 0% without any treatments, but increased to 1.7% under four combined treatments (clean and trim, watering, weeding, and cover with plastic to retain moisture) after being seeded in the field for 8 weeks. Hybrid offspring appeared to possess segregated traits following artificially controlled pollination. While hybrids of transgenic poplar and non-transgenic poplar can be excellent germplasm, gene flow should be monitored. Transgene expression in grafted scion and rootstock of transgenic poplar is reviewed. The transgenic poplar studied appears to be safe; no ecological or environmental harm has been observed in China.
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Abe, Tomoko, Hiroyuki Ichida, Yoriko Hayashi, et al. "Ion beam mutagenesis - an innovative and effective method for plant breeding and gene discovery." In Mutation breeding, genetic diversity and crop adaptation to climate change. CABI, 2021. http://dx.doi.org/10.1079/9781789249095.0042.
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Abstract We have developed a unique technology for mutation induction of plants using energetic ion beams at the RI Beam Factory (RIBF) of Rikagaku Kenkyūjo (RIKEN) (Institute of Physical and Chemical Research). Ion beams effectively induce mutations at relatively low doses without severely inhibiting growth. The irradiation treatment can be given to various plant materials and mutation can be induced in a short time, between seconds and a few minutes. The linear energy transfer (LET) of ions depends on the nuclide and velocity. Since LET value affects the mutation frequency, it is an important parameter to determine the most effective irradiation condition in mutagenesis. We determined the most effective dose in each LET for mutation induction in imbibed rice seeds. Subsequently, we analysed the mutated DNA responsible for the phenotype in morphological mutants. Most of the mutations were small deletions of less than 100 bp. Irradiations of C-ions and Ne-ions are effective for plant breeding because of the very high mutation rate and sufficient energy to disrupt a single gene. On the other hand, all mutations induced by Ar-ion (290 keV/μm) irradiation were large deletions ranging from 176 bp to approximately 620 kb. The average number of mutations in the target exon regions was 7.3, 8.5 and 4.3 per M<sub>3</sub> mutant plant in C-ions, Ne-ions and Ar-ions, respectively. The number of mutations induced by heavy-ion irradiation was relatively small. We could identify six responsible genes for eight mutants induced by C-ion and Ne-ion irradiations and two responsible genes for four mutants induced by Ar-ion irradiation. Three of these were genes not previously described.
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Garanto, Alejandro. "Delivery of Antisense Oligonucleotides to the Mouse Retina." In Methods in Molecular Biology. Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2010-6_22.
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AbstractThe eye is the organ in charge of vision and, given its properties, has become an excellent organ to test genetic therapies, including antisense oligonucleotide (AON) technology. In fact, the first AON receiving FDA and EMA approval was meant to treat an eye condition. Currently, dozens of clinical trials are being conducted for a variety of subtypes of inherited retinal disease. Although most of them are based on gene augmentation therapies, a phase 3 and two phase 1/2 clinical trials using AONs are ongoing. Since the retina is a layered structure of nondividing cells, obtaining human retinal tissue and expanding it in the lab is not possible, unless induced pluripotent stem cell technology is used. Mouse models have helped to elucidate the function of many genes, and the retinal structure is quite similar to that of humans. Thus, drug delivery to the mouse eye can provide valuable information for further optimization of therapies. In this chapter, the protocol for intravitreal injections of AONs is described in detail.
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Paro, Renato, Ueli Grossniklaus, Raffaella Santoro, and Anton Wutz. "Biology of Chromatin." In Introduction to Epigenetics. Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-68670-3_1.
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AbstractThis chapter provides an introduction to chromatin. We will examine the organization of the genome into a nucleosomal structure. DNA is wrapped around a globular complex of 8 core histone proteins, two of each histone H2A, H2B, H3, and H4. This nucleosomal arrangement is the context in which information can be established along the sequence of the DNA for regulating different aspects of the chromosome, including transcription, DNA replication and repair processes, recombination, kinetochore function, and telomere function. Posttranslational modifications of histone proteins and modifications of DNA bases underlie chromatin-based epigenetic regulation. Enzymes that catalyze histone modifications are considered writers. Conceptually, erasers remove these modifications, and readers are proteins binding these modifications and can target specific functions. On a larger scale, the 3-dimensional (3D) organization of chromatin in the nucleus also contributes to gene regulation. Whereas chromosomes are condensed during mitosis and segregated during cell division, they occupy discrete volumes called chromosome territories during interphase. Looping or folding of DNA can bring regulatory elements including enhancers close to gene promoters. Recent techniques facilitate understanding of 3D contacts at high resolution. Lastly, chromatin is dynamic and changes in histone occupancy, histone modifications, and accessibility of DNA contribute to epigenetic regulation.
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Maroni, Gustavo. "Size Variations Among the Elements that Constitute the Genes of Drosophila (Leader, coding region 3’ Untranslated Region, Exons, Introns)." In An Atlas of Drosophila Genes. Oxford University PressNew York, NY, 1993. http://dx.doi.org/10.1093/oso/9780195071160.003.0034.
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Abstract The discussion in this chapter centers around two questions: (1) what are the size ranges of the various elements that constitute a functional gene? And (2) is there a correlation between the size of one element and the size of another. The data analyzed in this chapter are derived from 73 of the genes presented in Part I. Because 12 of those 73 genes have multiple transcripts, they encompass a total of 87 transcripts. Two partly overlapping datasets can be examined: Dataset A includes all 87 transcripts, but elements shared by different transcripts of the same gene are considered only once. For example, if two transcripts of a gene differ only with respect to the poly (A) site, both 3’ untranslated regions (3’ UTR) are included in the analysis, but the leader is counted only once, since it is the same for both transcripts. Dataset B includes only one representative from each family of related genes or from the group of multiple transcripts of a given gene. In this case the sample is reduced to 40 “unrelated” transcripts. The size of a few elements were found to be outside the expected size range suggested by statistical analyses and so were excluded from the analysis. These elements are the 3’ UTR ofbsg25D II and the leader, 3’ UTR and introns of Ubx.
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Kuipers, Anja G. J., Evert Jacobsen,, and Richard G. F. Visser. "Applications of Antisense Technology in Plants." In Antisense Technology. Oxford University PressOxford, 1997. http://dx.doi.org/10.1093/oso/9780199635832.003.0009.
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Abstract In 1984, Izant and Weintraub (1) published the first experiments on the antisense effect of gene constructs in eukaryotic cells. Since then, the inhibition of gene expression via antisense RNA has developed into a technique that is widely applied in molecular biology. In the field of plant biotechnology and plant molecular biology Ecker and Davis (2) were the first to describe a transient assay system in which carrot protoplasts were simultaneously electro-porated with sense and antisense CAT genes. This resulted in the inhibition of CAT activity of up to 95% as compared to the control which only contained a sense CAT gene. These initial studies were followed by experiments with various plant species, in which the expression of a wide variety of genes has been effectively inhibited, either transiently with in vitro transcribed antisense RNAs, or by the introduction of antisense genes via transformation (reviewed in refs 3-5).
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Reddy, P. Sanjeeva, and Brent H. Cochran. "Novel gene transcription in response to growth factors: gene isolation by differential plaque hybridization." In Growth Factors. Oxford University PressOxford, 1993. http://dx.doi.org/10.1093/oso/9780199633609.003.0004.
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Abstract Differential cDNA cloning can be used to identify differences in gene expression between any two populations of cells. This method has been used in a variety of experimental situations, but has been especially successful in allowing for the isolation of genes which are regulated by growth factors. These screens have identified genes which are regulated by PDGF (1), TPA (2), IL-2 (3), serum (4, 5), and NGF (6) to name only a few. This analysis has increased our knowledge not only about the types of genes which are regulated by different growth factors, but also helped identify entirely new gene families such as the bZIP proteins and novel cytokines (7, 8). These growth factor regulated genes are often referred to as immediate early genes, or early response genes since many of them can be induced by growth factors in the absence of new protein synthesis. For review, see Herschman, 1991 (9).
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Beatty, Barbara G., and Stephen W. Scherer. "Human chromosome mapping of single copy genes." In Fish. Oxford University PressOxford, 2002. http://dx.doi.org/10.1093/oso/9780199638833.003.0003.
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Abstract One of the most common applications of FISH is the localization of single copy genes to a specific chromosomal band on metaphase chromosomes (1-3). Identifying the sub chromosomal position of a gene in both normal and disease states can provide valuable information regarding its biological and/or clinical significance. In some situations, mapping a newly identified gene to regions of chromosomal deletion or amplification, to translocation breakpoints, or to a region near a gene known to be involved in a particular disease process (4-6) can supply essential clues which may lead to important clinical applications (see Chapter 9).
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Holland, Peter W. H. "Cloning Genes Using the Polymerase Chain Reaction." In Essential Developmental Biology. Oxford University PressOxford, 1993. http://dx.doi.org/10.1093/oso/9780199634231.003.0025.
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Abstract The polymerase chain reaction (PCR) is an in vitro method for amplifying regions of DNA, utilizing information based on nucleotide sequences flanking the region of interest (1-3). PCR is used in the study of DNA sequence variation, genomic organization or gene expression (the latter using cDNA as a template, reverse transcribed from RNA; Chapter 24). PCR can also be used to amplify DNA from previously uncharacterized genes, if predictions can be made regarding the sequence of part of the gene(s).
Konferenzberichte zum Thema "CAV-3 gene"
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K. Alkhudhairy, Miaad, та Elhassan Benyagoub. "Frequency of Genes Mediated β-lactams Resistance in Acinetobacter Baumannii Isolates from Iraq". У X INTERNATIONAL CONGRESS OF PURE AND APPLIED TECHNOLOGICAL SCIENCES. Rimar Academy, 2023. http://dx.doi.org/10.47832/minarcongress10-2.
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Background: Acinetobacter baumannii is a nosocomial virulent microorganism that can cause acute and chronic infections in burn patients. The aim of the study: Diagnosis of genes mediated β-lactams resistance among test isolates. Materials and Methods: 649 swabs collected from inpatients with burn-wound infections at a burn center in Al-Najaf Province/ Iraq, from August 2022 to February 2023. Results: 68/ 649 (10.5%) isolates of Acinetobacter baumannii were identified according to microscopically, cultural, and biochemical features. 22 (32.4%) isolates were found able to produce extended-spectrum β-lactamases by using the double disks synergy method, and these producers tested by polymerase chain reactions technique for molecular determination β-lactams resistance encoding genes. This technique determined that the frequency of a single blaTEM gene and a single blaCTXM gene was 3/ 22 (13.6%) for each one among the test isolates and that 9/ 22 (41%) isolates possessed linked genes: the blaCTXM and blaTEM genes, whereas the blaSHV gene was not identified in any test isolate. Conclusions: The co-associated (blaTEM and blaCTXM) genes were revealed to be prevalent among the test isolates
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Olton, Dana, Dong Hyun Lee, Charles Sfeir, and Prashant N. Kumta. "Novel Nanostructured Calcium Phosphate Based Delivery Systems for Non-Viral Gene Delivery." In ASME 2007 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2007. http://dx.doi.org/10.1115/sbc2007-176286.
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Calcium phosphate (CaP) based approaches remain an attractive option for delivering plasmid DNA (pDNA) into cultured cells [1, 2]. However, two major limitations associated with this vector exist. First, it yields lower transfection efficiencies with respect to its’ viral counterparts. Second, CaP mediated gene delivery leads to transient transgene expression. Thus, we hypothesized that these concerns could respectively be addressed by: (1) synthesizing particles with precise control of the materials’ parameters including (i.e. Ca/P ratio, particle size, crystal structure, and microstructure) and (2) incorporating the particles into a 3-D biodegradable fibrin scaffold. The goal of this study was therefore to synthesize and optimize the efficacy of both nano-structured CaP (NanoCaPs) particles and a composite scaffold comprised of fibrin, CaP and pDNA for non-viral gene delivery applications.
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Chakraborty, Amlan, Venkatakrishna R. Jala, Sutirtha Chakraborty, R. Eric Berson, M. Keith Sharp, and Bodduluri Haribabu. "Bidirectional Oscillatory Shear Stress Increases Pro-Atherogenic Gene Expressions (I-CAM1, E-Selectin and IL-6) in Endothelial Cells." In ASME 2011 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2011. http://dx.doi.org/10.1115/sbc2011-53747.
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Wall shear stress (WSS) plays a key role in altering intracellular pathways and gene expression of endothelial cells, and has significant impacts on atherosclerotic plaque development (1–3). Further, the atherogenic regulators Leukotriene B4 (LTB4) and Lipopolysaccharide (LPS) have significant impacts on the pathophysiology of many inflammatory diseases. This study investigates the effects of oscillatory shear directionality on pro-atherogenic gene expression (I-CAM, E-Selectin, and IL-6) in the presence of LTB4 and LPS. An orbital shaker was used to expose the endothelial cells to oscillatory shear in culture dishes, and Computational fluid dynamics (CFD) was applied to quantify the shear stress on the bottom of the orbiting dish. Directionality of oscillatory shear was characterized by a newly developed hemodynamic parameter — Directional oscillatory shear index (DOSI), which was demonstrated in a previous study to significantly impact cell morphology (4). Results showed that DOSI significantly altered gene expression. Therefore, directionality of shear modulates atherosclerotic gene expression in vitro and thus, may influence the formation of atherosclerotic plaque in vivo.
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Vaughan, Andrew T., Rebecca Wright, and Katrina Slemmons. "Abstract B35: Estradiol drives MLL gene fusions in infant acute leukemia." In Abstracts: AACR Special Conference: Pediatric Cancer at the Crossroads: Translating Discovery into Improved Outcomes; November 3-6, 2013; San Diego, CA. American Association for Cancer Research, 2014. http://dx.doi.org/10.1158/1538-7445.pedcan-b35.
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Bernaedi, F., V. Bertagnolo, S. Bartolai, L. Rossi, F. Panicucci, and F. Conconi. "A POINT MUTATION AND A GENE DELETION OF FVIII GENE IN SEVERE HAEMOPHILIA." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644047.
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The presence of Factor VIII (FVIII) gene lesions has been investigated in 100 haemophilia A patients using cDNA probes for the 3'part of FVIII gene (exons 14-26 ).In two related severe patients without inhibitor a deletion removesthe exon 26; the gene lesion has been confirmed with several restriction enzymes and has been shown by densitometry of the autoradiographic pattern in a woman of the same family. The complete deletionof the exon 26 has been described by Gitschier et al. in a patient with inhibitor. Thus the comparison of the end points of the two deletions could help to define the mechanism originating these gene lesions and the relation between gene lesions and the presence of antibody.In a patient with severe Haemophilia and without inhibitor a mutation removing the TaqI site in the exon 24 and originating an abnormal band of 4.2 Kb has been found. A C→T transition in this TaqI site, originating a nonsense codon and a new Hindlll site, has been reported by Gitschier et al in a patient presenting inhibitor. The DNA from our patient tested with Hindlll shows a normal pattern thus indicating a C→T transition in the antisense strand. This mutation should causean aminoacid change (CGA→CAA, Arg→Gln) possiblyresponsible for the FVIII inactivation but that does not remove theantigenic determinants present in the COOH terminal part of FVIII.In addition the same mutation has been observed in an unrelated (asdemonstrated by RFLPs analysis) Italian haemophilic patient confirming the observation of Youssoufian et al that TaqI sites are mutational hot spots in FVIII gene.
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Birch, David G., Gabriel H. Travis, Kirsten G. Locke, and Donald C. Hood. "Rod Ergs in Mice and Humans with Putative Null Mutations in the RDS Gene." In Vision Science and its Applications. Optica Publishing Group, 1997. http://dx.doi.org/10.1364/vsia.1997.ma.4.
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Mutations causing autosomal dominant retinitis pigmentosa (RP) have been identified in RDS, the human homologue of the gene that was first identified and isolated as the cause of mouse "retinal degeneration slow" or rds (1, 2). The rds gene encodes rds/peripherin, an integral membrane glycoprotein located in outer segment disks (1, 3, 4). More than 15 distinct disease-causing mutations in the RDS gene have been reported (5). The clinical phenotypes include adRP, dominant retinitis punctata albescens, dominant butterfly shaped pigment dystrophy of the fovea and autosomal dominant macular degeneration (6). That RDS mutation can cause either RP and/or macular degeneration is consistent with the observation that the protein is expressed in both rods and cones, though its exact functional role in each photoreceptor must be different. Finally, there is an additional form of retinitis pigmentosa, digenic RP (7) that results from a combination of one mutation in RDS and one in R0M1. Neither mutation alone in a heterozygote causes degeneration.
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Bhardwaj, Anjana, Nivetha Ganesan, Constance T. Albarracin, and Isabelle Bedrosian. "Abstract A115: Annexin A1: A novel gene target of miRNA-21." In Abstracts: AACR Special Conference on Advances in Breast Cancer Research: Genetics, Biology, and Clinical Applications - October 3-6, 2013; San Diego, CA. American Association for Cancer Research, 2013. http://dx.doi.org/10.1158/1557-3125.advbc-a115.
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Badia, Bruno de Mattos Lombardi, Roberta Ismael Lacerda Machado, Wladimir Bocca Vieira de Rezende Pinto, et al. "Blurred Lines – Is the distinction between CIDP and CMT always clear?" In XIII Congresso Paulista de Neurologia. Zeppelini Editorial e Comunicação, 2021. http://dx.doi.org/10.5327/1516-3180.015.
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Introduction: Charcot-Marie-Tooth disease (CMT) is a group of inherited sensorymotor neuropathies with variable age of onset, clinical and neurophysiological patterns but often with a chronic slow progression. Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is an immune-mediated neuropathy with a relapsing clinical course and typically good response to corticosteroids or other therapies. The distinction between these two conditions can be made with aid of clinical history, neurophysiological studies and genetic testing in the vast majority of cases. However, an overlap between them can occur. Methods: We describe four Brazilian patients under clinical follow-up at our service with genetic diagnosis of CMT and clinical and neurophysiological features compatible with a concurrent CIDP diagnosis. Results: Four cases of different CMT subtypes with co-occurrence of an immunemediated neuropathy compatible with CIDP were reported. The patients were all unrelated, two males and two females, age range from 3 to 45 years. The genetic mutations were the following: hemizygous pathogenic variant c.514C>T (p.Pro172Ser) in GJB1 gene (CMT1X), duplication of PMP22 gene (CMT1A), simple heterozygous pathogenic variant c.188_190delCCT (p.Ser64del) in MPZ gene (CMT1B) and homozygous pathogenic variant c.122T>C (p.Ile41Thr) in FIG4 gene (CMT4J). All four patients presented with relapsing or subacute worsening of neurological symptoms, demyelinating non-uniform features in neurophysiological studies including conduction blocks and elevated cerebrospinal fluid (CSF) protein levels without pleocytosis. Three patients (3/4) improved after treatment with corticosteroids, immunoglobulin or cyclophosphamide with variable clinical response. Conclusion: CMT and CIDP are different conditions involving the peripheral nervous system and the distinction between them usually is possible with the appropriate assessment. The overlap between them is possible and we report four cases with this association.
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O'hara, Patrick J., Frank A. Grant, A. Betty, J. Haldmen, and Mark J. Murray. "Structure of the Human Factor VII Gene." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643786.
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Factor VII is a member of a family of vitamin K-dependent, gamma-carboxylated plasma protein which includes factor IX, factor X, protein C, protein S and prothrombin. Activated factor VII (factor Vila) is a plasma serine protease which participates in a cascade of reactions leading to the coagulation of blood. Two overlapping genomic clones containing sequences encoding human factor VII were isolated and characterized. The complete sequence of the gene was determined and found to span 12.8 kilobases. The mRNA for factor VII as demonstrated by cDNA cloning is polyadenylated at multiple sites but contains only one AAUAAA poly-A signal sequence. The mRNA can undergo alternative splicing forming one transcript containing eight segments as exons and another with an additional exon which encodes a larger pre-pro leader sequence. The portion of the pre-pro leader coded for by the additional exon has no known counterpart in the other vitamin K-dependent proteins. The positions of the introns with respect to the amino acid sequence encoded by the eight essential exons of factor VII are the same as those present in factor IX, factor X, protein C and the first three exons of prothrombin. These exons code for domains generally conserved among members of this gene family, including a pre-pro leader (the essential exon la and alternative exon lb), a gamma-carboxylated domain (exons 2 and 3) a growth factor domain (exons 4 and 5) an activation region (exon 6) and a serine protease (exon 8). The corresponding introns in these genes are dissimilar with respect to size and sequence, with the exception of the third intron in factor VII and protein C. Four introns and a portion of exon 8 in factor VII contain regions made up of tandem repeats of oligonucleotide monomer elements. More than a quarter of the intron sequences and more than a third of the 3' untranslated portion of the mRNA transcript consist of these minisatellite tandem repeats. This type of structure is responsible for polymorphisms due to allelic variation in repeat copy number in other areas of the human genome. Tandem repeats can evolve as a result of random crossover in DNA whose sequence is not maintained by selection. This suggests that much of the sequence information present in the introns and untranslated portion of the message is dispensable.
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Fleming, Paul, and Tara Dalton. "One-Step Reverse-Transcription PCR on a High-Throughput Micro-Fluidic Device." In ASME 2009 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2009. http://dx.doi.org/10.1115/sbc2009-206623.
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One step reverse-transcription polymerase chain reaction (RT-PCR) assays are an attractive option for further automating gene detection assays. One-step assays can reduce hands–on-time and the risk of sample crossover and contamination. The one-step chemistries are showing increasing use in virus detection and have been reported, in some cases, to be more appropriate than their two-step counterparts [1, 2]. Previous work presented by the Stokes Institute research group outlined a micro fluidic based continuous flow instrument which performed high throughput qPCR in nanolitre sized droplets [3]. This instrument had advantages over commercially available instruments in that it could process far more than the traditional 96 or 384 reaction setup in a single run and the reaction volume was reduced from 20–50 μl down to 30–100 nl sized droplets. Combining one-step chemistry with the technology offered by the devices being developed would lead to a high-throughput RNA-to-signal system capable of reverse transcribing and performing PCR on thousands of nanolitre sized reactions every day. It is envisaged that this technology will also lead to gene expression from single cells contained in nanolitre sized droplets. In this paper, a study was conducted in which an extra thermal region, manufactured from aluminium, was added to the existing continuous flow instruments. This region was maintained at a temperature suitable for reverse transcription, which was 48°C for the one-step kit tested. The thermal region was also a suitable length to maintain the sample at the required temperature for 15 minutes. Using a commercially available one step RT-PCR kit (TaqMan® RNA-to-CT™ 1-Step Kit, 4392653), the device was evaluated for its potential to perform one-step RT-PCR in continuously flowing nanolitre sized droplets. Electrophoresis gels were initially used in assessing specific amplification before an end-point detection method was utilized. RNA was extracted from the leukemic REH cell line with the housekeeping gene, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) as the gene of interest. To investigate the possibility of further reducing sample preparation and facilitating further automation, amplification from cell lysates without nucleic acid extraction was carried out on the device. Cell lysates were prepared using the cell lysis buffer from the TaqMan® Gene Expression Cells-to-CT™ Kit (Cat #AM1728). It was found that the device was successful in one-step RT-PCR from extracted RNA samples and samples from cell lysates without nucleic acid extraction.
Berichte der Organisationen zum Thema "CAV-3 gene"
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Levin, Ilan, John W. Scott, Moshe Lapidot, and Moshe Reuveni. Fine mapping, functional analysis and pyramiding of genes controlling begomovirus resistance in tomato. United States Department of Agriculture, 2014. http://dx.doi.org/10.32747/2014.7594406.bard.
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Abstract. Tomato yellow leaf curl virus (TYLCV), a monopartitebegomovirus, is one of the most devastating viruses of cultivated tomatoes and poses increasing threat to tomato production worldwide. Because all accessions of the cultivated tomato are susceptible to these viruses, wild tomato species have become a valuable resource of resistance genes. QTL controlling resistance to TYLCV and other begomoviruses (Ty loci) were introgressed from several wild tomato species and mapped to the tomato genome. Additionally, a non-isogenic F₁diallel study demonstrated that several of these resistance sources may interact with each other, and in some cases generate hybrid plants displaying lower symptoms and higher fruit yield compared to their parental lines, while their respective resistance genes are not necessarily allelic. This suggests that pyramiding genes originating from different resistance sources can be effective in obtaining lines and cultivars which are highly resistant to begomoviruses. Molecular tools needed to test this hypothesis have been developed by our labs and can thus significantly improve our understanding of the mechanisms of begomovirus resistance and how to efficiently exploit them to develop wider and more durable resistance. Five non-allelic Ty loci with relatively major effects have been mapped to the tomato genome using molecular DNA markers, thereby establishing tools for efficient marker assisted selection, pyramiding of multiple genes, and map based gene cloning: Ty-1, Ty-2, Ty-3, Ty-4, and ty-5. This research focused on Ty-3 and Ty-4 due to their broad range of resistance to different begomoviruses, including ToMoV, and on ty-5 due to its exceptionally high level of resistance to TYLCV and other begomoviruses. Our aims were: (1) clone Ty-3, and fine map Ty-4 and Ty-5 genes, (2)introgress each gene into two backgroundsand develop semi isogenic lines harboring all possible combinations of the three genes while minimizing linkage-drag, (3) test the resulting lines, and F₁ hybrids made with them, for symptom severity and yield components, and (4) identify and functionally characterize candidate genes that map to chromosomal segments which harbor the resistance loci. During the course of this research we have: (1) found that the allelic Ty-1 and Ty-3 represent two alternative alleles of the gene coding DFDGD-RDRP; (2) found that ty-5is highly likely encoded by the messenger RNA surveillance factor PELOTA (validation is at progress with positive results); (3) continued the map-based cloning of Ty-4; (4) generated all possible gene combinations among Ty-1, Ty-3 and ty-5, including their F₁ counterparts, and tested them for TYLCV and ToMoV resistance; (5) found that the symptomless line TY172, carrying ty-5, also carries a novel allele of Ty-1 (termed Ty-1ⱽ). The main scientific and agricultural implications of this research are as follows: (1) We have developed recombination free DNA markers that will substantially facilitate the introgression of Ty-1, Ty-3 and ty-5 as well as their combinations; (2) We have identified the genes controlling TYLCV resistance at the Ty-1/Ty-3 and ty-5 loci, thus enabling an in-depth analyses of the mechanisms that facilitate begomovirus resistance; (3) Pyramiding of Ty resistance loci is highly effective in providing significantly higher TYLCV resistance.
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Reisch, Bruce, Avichai Perl, Julie Kikkert, Ruth Ben-Arie, and Rachel Gollop. Use of Anti-Fungal Gene Synergisms for Improved Foliar and Fruit Disease Tolerance in Transgenic Grapes. United States Department of Agriculture, 2002. http://dx.doi.org/10.32747/2002.7575292.bard.
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Original objectives . 1. Test anti-fungal gene products for activity against Uncinula necator, Aspergillus niger, Rhizopus stolonifer and Botrytis cinerea. 2. For Agrobacterium transformation, design appropriate vectors with gene combinations. 3. Use biolistic bombardment and Agrobacterium for transformation of important cultivars. 4. Characterize gene expression in transformants, as well as level of powdery mildew and Botrytis resistance in foliage of transformed plants. Background The production of new grape cultivars by conventional breeding is a complex and time-consuming process. Transferring individual traits via single genes into elite cultivars was proposed as a viable strategy, especially for vegetatively propagated crops such as grapevines. The availability of effective genetic transformation procedures, the existence of genes able to reduce pathogen stress, and improved in vitro culture methods for grapes, were combined to serve the objective of this proposal. Effective deployment of resistance genes would reduce production costs and increase crop quality, and several such genes and combinations were used in this project. Progress The efficacy of two-way combinations of Trichoderma endochitinase (CHIT42), synthetic peptide ESF12 and resveratrol upon the control of growth of Botrytis cinerea and Penicillium digitatum were evaluated in vitro. All pairwise interactions were additive but not synergistic. Per objective 2, suitable vectors with important gene combinations for Agrobacterium transformation were designed. In addition, multiple gene co-transformation by particle bombardment was also tested successfully. In New York, transformation work focused on cultivars Chardonnay and Merlot, while the technology in Israel was extended to 41B, R. 110, Prime, Italia, Gamay, Chardonnay and Velika. Transgenic plant production is summarized in the appendix. Among plants developed in Israel, endochitinase expression was assayed via the MuchT assay using material just 1-5 days after co-cultivation. Plants of cv. Sugraone carrying the gene coding for ESF12, a short anti-fungal lytic peptide under the control of the double 358 promoter, were produced. Leaf extracts of two plants showed inhibition zones that developed within 48 h indicating the inhibitory effect of the leaf extracts on the six species of bacteria. X fastidiosa, the causal organism of Pierce's disease, was very sensitive to leaf extracts from ESF12 transformed plants. Further work is needed to verify the agricultural utility of ESF12 transformants. In New York, some transformants were resistant to powdery mildew and Botrytis fruit rot. Major conclusions, solutions, achievements and implications The following scientific achievements resulted from this cooperative BARD project: 1. Development and improvement of embryogenesis and tissue culture manipulation in grape, while extending these procedures to several agriculturally important cultivars both in Israel and USA. 2. Development and improvement of novel transformation procedures while developing transformation techniques for grape and other recalcitrant species. 3. Production of transgenic grapevines, characterization of transformed vines while studying the expression patterns of a marker gene under the control of different promoter as the 35S CaMV in different part of the plants including flowers and fruits. 4. Expression of anti-fungal genes in grape: establishment of transgenic plants and evaluation of gene expression. Development of techniques to insert multiple genes. 5. Isolation of novel grape specific promoter to control the expression of future antimicrobial genes. It is of great importance to report that significant progress was made in not only the development of transgenic grapevines, but also in the evaluation of their potential for increased resistance to disease as compared with the non engineered cultivar. In several cases, increased disease resistance was observed. More research and development is still needed before a product can be commercialized, yet our project lays a framework for further investigations.
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Palukaitis, Peter, Amit Gal-On, Milton Zaitlin, and Victor Gaba. Virus Synergy in Transgenic Plants. United States Department of Agriculture, 2000. http://dx.doi.org/10.32747/2000.7573074.bard.
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Transgenic plants expressing viral genes offer novel means of engendering resistance to those viruses. However, some viruses interact synergistically with other viruses and it is now known that transgenic plants expressing particular genes of one virus may also mediate synergy with a second virus. Thus, our specific objectives were to (1) determine if transgenic plants resistant to one virus showed synergy with another virus; (2) determine what viral sequences were essential for synergy; and (3) determine whether one of more mechanisms were involved i synergy. This project would also enable an evaluation of the risks of synergism associated with the use of such transgenic plants. The conclusion deriving from this project are as follows: - There is more than one mechanism of synergy. - The CMV 2b gene is required for synergistic interactions. - Synergy between a potyvirus and CMV can break natural resistance limiting CMV movement. - Synergy operates at two levels - increase in virus accumulation and increase in pathology - independently of each other. - Various sequences of CMV can interact with the host to alter pathogenicity and affect virus accumulation. - The effect of synergy on CMV satellite RNA accumulatio varies in different systems. - The HC-Pro gene may only function in host plant species to induce synergy. - The HC-Pro is a host range determinant of potyviruses. - Transgenic plants expressing some viral sequences showed synergy with one or more viruses. Transgenic plants expressing CMV RNA 1, PVY NIb and the TMV 30K gene all showed synergy with at least one unrelated virus. - Transgenic plants expressing some viral sequences showed interference with the infection of unrelated viruses. Transgenic plants expressing the TMV 30K, 54K and 126K genes, the PVY NIb gene, or the CMV 3a gene all showed some level of interference with the accumulation (and in some cases the pathology) of unrelated viruses. From our observations, there are agricultural implications to the above conclusions. It is apparent that before they are released commercially, transgenic plants expressing viral sequences for resistance to one virus need to be evaluated fro two properties: - Synergism to unrelated viruses that infect the same plant. Most of these evaluations can be made in the greenhouse, and many can be predicted from the known literature of viruses known to interact with each other. In other cases, where transgenic plants are being generated from new plant species, the main corresponding viruses from the same known interacting genera (e.g., potexviruses and cucumoviruses, potyviruses and cucumoviruses, tobamoviruses and potexviruses, etc.) should be evaluated. - Inhibition or enhancement of other resistance genes. Although it is unlikely that plants to be released would be transformed with HC-Pro or 2b genes, there may be other viral genes that can affect the expression of plant genes encoding resistance to other pathogens. Therefore, transgenic plants expressing viral genes to engender pathogen-derived resistance should be evaluated against a spectrum of other pathogens, to determine whether those resistance activities are still present, have been lost, or have been enhanced!
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Grumet, Rebecca, Rafael Perl-Treves, and Jack Staub. Ethylene Mediated Regulation of Cucumis Reproduction - from Sex Expression to Fruit Set. United States Department of Agriculture, 2010. http://dx.doi.org/10.32747/2010.7696533.bard.
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Reproductive development is a critical determinant of agricultural yield. For species with unisexual flowers, floral secualdifferentation adds additional complexity, that can influenec productivity. The hormone ethylene has long, been known to play a primary role in sex determination in the Cucumis species cucumber (C. sativus) and melon (C. melo). Our objectives were to: (1) Determine critical sites of ethylene production and perception for sex determination; (2) Identify additional ethylene related genes associated with sex expression; and (3) Examine the role of environment ami prior fruit set on sex expression, pistillate flower maturation, and fruit set. We made progress in each of these areas. (1) Transgenic melon produced with the Arabidopsis dominant negative ethylene perception mutant gene, etrl-1, under the control of floral primordia targeted promoters [AP3 (petal and stamen) and CRC (carpel and nectary)], showed that ethylene perception by the stamen primordia, rather than carpel primordia, is critical for carpel development at the time of sex determination. Transgenic melons also were produced with the ethylene production enzyme gene. ACS, encoding l-aminocyclopropane-lcarboylate synthase, fused to the AP3 or CRC promoters. Consistent with the etr1-1 results, CRC::ACS did not increase femaleness; however, AP3::ACS reduced or eliminated male flower production. The effects of AP3:ACS were stronger than those of 35S::ACS plants, demonstratin g the importance of targeted expression, while avoiding disadvantages of constitutive ethylene production. (2) Linkage analysis coupled with SNP discovery was per formed on ethylene and floral development genes in cucumber populations segregating for the three major sex genes. A break-through towards cloning the cucumber M gene occurred when the melon andromonoecious gene (a), an ACS gene, was cloned in 2008. Both cucumber M and melon a suppress stamen development in pistillate flowers. We hypothesized that cucumber M could be orthologous to melon a, and found that mutations in CsACS2 co-segregated perfectly with the M gene. We also sought to identify miRNA molecules associated with sex determination. miRNA159, whose target in Arabidopsis is GAMYB[a transcription factor gene mediating response to10 gibberellin (GA)], was more highly expressed in young female buds than male. Since GA promotes maleness in cucumber, a micro RNA that counteracts GAMYB could promote femaleness. miRNA157, which in other plants targets transcription factors involved in flower development , was expressed in young male buds and mature flower anthers. (3) Gene expression profiling showed that ethylene-, senescence-, stress- and ubiquitin-related genes were up-regulated in senescing and inhibited fruits, while those undergoing successful fruit set up-regulated photosynthesis, respiration and metabolic genes. Melon plants can change sex expression in response to environmental conditions, leading to changes in yield potential. Unique melon lines with varying sex expression were developed and evaluated in the field in Hancock, Wisconsin . Environmental changes during the growing season influenced sex expression in highly inbred melon lines. Collectively these results are of significance for understanding regulation of sex expression. The fact that both cucumber sex loci identified so far (F and M) encode isoforms of the same ethylene synthesis enzyme, underscores the importance of ethylene as the main sex determining hormone in cucumber. The targeting studies give insight into developmental switch points and suggest a means to develop lines with earlier carpel-bearing flower production and fruit set. These results are of significance for understanding regulation of sex expression to facilitate shorter growing seasons and earlier time to market. Field results provide information for development of management strategies for commercial production of melon cultivars with different sex expression characteristics during fruit production.
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Xu, Jin-Rong, and Amir Sharon. Comparative studies of fungal pathogeneses in two hemibiotrophs: Magnaporthe grisea and Colletotrichum gloeosporioides. United States Department of Agriculture, 2008. http://dx.doi.org/10.32747/2008.7695585.bard.
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Plant pathogenic fungi have various life styles and different plant infection strategies. Hemibiotrophs like Magnaporthe grisea and Colletotrichum species develop specialized structures during plant infection. The goal of this study was to identify, characterize, and compare genes required for plant infection in M. grisea and C. gloeosporioides. Specific objectives are to: 1) further characterize genes identified in the preliminary studies of C. gloeosporioides and M. grisea;2) identify and characterize additional fungal genes tagged by GFP; and 3) identify in planta growth and appressorium-specific genes by subtractive hybridization and transcript profiling by the LongSAGE method. In this study, the PI and Co-PI collaborated closely on studies in M. grisea and C. gloeosporioides. In M. grisea, REMI and ATMT were used to transform the wildtype with promoter-less EGFP constructs. A total of 28 mutants defective in different plant infection processes or expressing EGFP during plant infection were identified. Genes disrupted in five selected mutants have been identified, including MG03295 that encodes a putative Rho GTPase. In transformant L1320, the transforming vector was inserted in the MIRI gene that encodes a nuclear protein. The expression of MIRI was highly induced during infection. Deletion and site-directed mutagenesis analyses were used to identify the promoter regions and elements that were essential for induced in planta expression of MIRI. This was the first detailed characterization of the promoter of an in planta gene in M. grisea and the MIRI promoter can be used to monitor infectious growth. In addition, the Agilent whole-genome array of M. grisea was used for microarray analyses with RNA samples from appressoria formed by the wild-type shain and the pmkl and mstl2 mutants. Over 200 genes were downregulated in the mst I 2 and pmkl mutants. Some of them are putative transcription factors that may regulate appressorium formation and infectious hyphal growth. In C. gloeosporioides, various REMI mutants showing different pathogenic behavior were identified and characterized. Mutants N3736 had a single insertion and was hyper-virulent. The gene disrupted in mutant3736 (named CgFMOI) encodes a FAD-dependent monooxygenase. Expression analyses linked the expression of the CgFMOI gene with the necrotrophic phase of fungal infection, and also suggest that expression of CgFMOl is unnecessary for the first stages of infection and for biotrophy establishment. All CgFMOl-silenced mutants had reduced virulence. In REMI mutant N159, the tagged gene encodes a putative copper transporter that is homologue of S. cerevisiae CTR2. In yeast, Ctr2 is a vacuolar transporter for moving copper from the vacuole to the cytoplasm. The gene was therefore termed CgCTR2. In addition to characterization of CgCTR2, we also conducted comparative analyses in M. grisea. The M. grisea CgCTR-2 homolog was isolated, knockout strains were generated and characterized and the M. grisea was used to complement the Nl 59 C. gloeosporioides mutant. Overall, we have accomplished most of proposed experiments and are in the process of organizing and publishing other data generated in this project. For objective 3, we used the microarray analysis approach. Several genes identified in this study are novel fungal virulence factors. They have the potential to be used as targets for developing more specific or effective fungicides. In the long run, comparative studies of fungal genes, such as our CgCTR2 work, may lead to better disease control strategies.
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Gal-On, Amit, Shou-Wei Ding, Victor P. Gaba, and Harry S. Paris. role of RNA-dependent RNA polymerase 1 in plant virus defense. United States Department of Agriculture, 2012. http://dx.doi.org/10.32747/2012.7597919.bard.
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Objectives: Our BARD proposal on the impact of RNA-dependent RNA polymerase 1 (RDR1) in plant defense against viruses was divided into four original objectives. 1. To examine whether a high level of dsRNA expression can stimulate RDR1 transcription independent of salicylic acid (SA) concentration. 2. To determine whether the high or low level of RDR1 transcript accumulation observed in virus resistant and susceptible cultivars is associated with viral resistance and susceptibility. 3. To define the biogenesis and function of RDR1-dependent endogenous siRNAs. 4. To understand why Cucumber mosaic virus (CMV) can overcome RDR1-dependent resistance. The objectives were slightly changed due to the unique finding that cucumber has four different RDR1 genes. Background to the topic: RDR1 is a key plant defense against viruses. RDR1 is induced by virus infection and produces viral and plant dsRNAs which are processed by DICERs to siRNAs. siRNAs guide specific viral and plant RNA cleavage or serve as primers for secondary amplification of viral-dsRNA by RDR. The proposal is based on our preliminary results that a. the association of siRNA and RDR1 accumulation with multiple virus resistance, and b. that virus infection induced the RDR1-dependent production of a new class of endogenous siRNAs. However, the precise mechanisms underlying RDR1 induction and siRNA biogenesis due to virus infection remain to be discovered in plants. Major conclusions, solutions and achievements: We found that in the cucurbit family (cucumber, melon, squash, watermelon) there are 3-4 RDR1 genes not documented in other plant families. This important finding required a change in the emphasis of our objectives. We characterized 4 RDR1s in cucumber and 3 in melon. We demonstrated that in cucumber RDR1b is apparently a new broad spectrum virus resistance gene, independent of SA. In melon RDR1b is truncated, and therefore is assumed to be the reason that melon is highly susceptible to many viruses. RDR1c is dramatically induced due to DNA and RNA virus infection, and inhibition of RDR1c expression led to increased virus accumulation which suggested its important on gene silencing/defense mechanism. We show that induction of antiviral RNAi in Arabidopsis is associated with production of a genetically distinct class of virus-activated siRNAs (vasiRNAs) by RNA dependent RNA polymerase-1 targeting hundreds of host genes for RNA silencing by Argonaute-2. Production of vasiRNAs is induced by viruses from two different super groups of RNA virus families, targeted for inhibition by CMV, and correlated with virus resistance independently of viral siRNAs. We propose that antiviral RNAi activate broad-spectrum antiviral activity via widespread silencing of host genes directed by vasiRNAs, in addition to specific antiviral defense Implications both scientific and agricultural: The RDR1b (resistance) gene can now be used as a transcription marker for broad virus resistance. The discovery of vasiRNAs expands the repertoire of siRNAs and suggests that the siRNA-processing activity of Dicer proteins may play a more important role in the regulation of plant and animal gene expression than is currently known. We assume that precise screening of the vasiRNA host targets will lead in the near future for identification of plant genes associate with virus diseases and perhaps other pathogens.
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Friedman, Haya, Julia Vrebalov, James Giovannoni, and Edna Pesis. Unravelling the Mode of Action of Ripening-Specific MADS-box Genes for Development of Tools to Improve Banana Fruit Shelf-life and Quality. United States Department of Agriculture, 2010. http://dx.doi.org/10.32747/2010.7592116.bard.
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Fruit deterioration is a consequence of a genetically-determined fruit ripening and senescence programs, in which developmental factors lead to a climacteric rise of ethylene production in ethylene-sensitive fruits such as tomato and banana. Breeding of tomato with extended fruit shelf life involves the incorporation of a mutation in RIN, a MADS-box transcription factor participating in developmental control signalling of ripening. The RIN mode of action is not fully understood, and it may be predicted to interact with other MADS-box genes to execute its effects. The overall goal of this study was to demonstrate conservation of ripening control functions between banana and tomato and thus, the potential to genetically extend shelf-life in banana based on tools developed in tomato. The specific objectives were: 1. To increase the collection of potential RIN-like genes from banana; 2. To verify their action as developmental regulators; 3. To elucidate MADS-box gene mode of action in ripening control; 4. To create transgenic banana plants that express low levels of endogenous Le-RIN- like, MaMADS- gene(s). We have conducted experiments in banana as well as in tomato. In tomato we have carried out the transformation of the tomato rin mutant with the MaMADS1 and MaMADS2 banana genes. We have also developed a number of domain swap constructs to functionally examine the ripening-specific aspects of the RIN gene. Our results show the RIN-C terminal region is essential for the gene to function in the ripening signalling pathway. We have further explored the tomato genome databases and recovered an additional MADS-box gene necessary for fruit ripening. This gene has been previously termed TAGL1 but has not been functionally characterized in transgenic plants. TAGL1 is induced during ripening and we have shown via RNAi repression that it is necessary for both fleshy fruit expansion and subsequent ripening. In banana we have cloned the full length of six MaMADS box genes from banana and determined their spatial and temporal expression patterns. We have created antibodies to MaMADS2 and initiated ChI assay. We have created four types of transgenic banana plants designed to reduce the levels of two of the MaMADS box genes. Our results show that the MaMADS-box genes expression in banana is dynamically changing after harvest and most of them are induced at the onset of the climacteric peak. Most likely, different MaMADS box genes are active in the pulp and peel and they are differently affected by ethylene. Only the MaMADS2 box gene expression is not affected by ethylene indicating that this gene might act upstream to the ethylene response pathway. The complementation analysis in tomato revealed that neither MaMADS1 nor MaMADS2 complement the rin mutation suggesting that they have functionally diverged sufficiently to not be able to interact in the context of the tomato ripening regulatory machinery. The developmental signalling pathways controlling ripening in banana and tomato are not identical and/or have diverged through evolution. Nevertheless, at least the genes MaMADS1 and MaMADS2 constitute part of the developmental control of ripening in banana, since transgenic banana plants with reduced levels of these genes are delayed in ripening. The detailed effect on peel and pulp, of these transgenic plants is underway. So far, these transgenic bananas can respond to exogenous ethylene, and they seem to ripen normally. The response to ethylene suggest that in banana the developmental pathway of ripening is different than that in tomato, because rin tomatoes do not ripen in response to exogenous ethylene, although they harbor the ethylene response capability This study has a major contribution both in scientific and agricultural aspects. Scientifically, it establishes the role of MaMADS box genes in a different crop-the banana. The developmental ripening pathway in banana is similar, but yet different from that of the model plant tomato and one of the major differences is related to ethylene effect on this pathway in banana. In addition, we have shown that different components of the MaMADS-box genes are employed in peel and pulp. The transgenic banana plants created can help to further study the ripening control in banana. An important and practical outcome of this project is that we have created several banana transgenic plants with fruit of extended shelf life. These bananas clearly demonstrate the potential of MaMADS gene control for extending shelf-life, enhancing fruit quality, increasing yield in export systems and for improving food security in areas where Musaspecies are staple food crops.
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Meir, Shimon, Michael Reid, Cai-Zhong Jiang, Amnon Lers, and Sonia Philosoph-Hadas. Molecular Studies of Postharvest Leaf and Flower Abscission. United States Department of Agriculture, 2005. http://dx.doi.org/10.32747/2005.7696523.bard.
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Original objectives: Understanding the regulation of abscission competence by exploring the nature and function of auxin-related gene expression changes in the leaf and pedicelAZs of tomato (as a model system), was the main goal of the previously submitted proposal. We proposed to achieve this goal by using microarray GeneChip analysis, to identify potential target genes for functional analysis by virus-induced gene silencing (VIGS). To increase the potential of accomplishing the objectives of the previously submitted proposal, we were asked by BARD to show feasibility for the use of these two modern techniques in our abscission system. Thus, the following new objectives were outlined for the one-year feasibility study: 1.to demonstrate the feasibility of the VIGS system in tomato to perform functional analysis of known abscission-related genes; 2. to demonstrate that by using microarray analysis we can identify target genes for further VIGS functional analysis. Background to the topic: It is a generally accepted model that auxin flux through the abscission zone (AZ) prevents organ abscission by rendering the AZ insensitive to ethylene. However, the molecular mechanisms responsible for acquisition of abscission competence and the way in which the auxin gradient modulates it are still unknown. Understanding this basic stage of the abscission process may provide us with future tools to control abscission for agricultural applications. Based on our previous study, performed to investigate the molecular changes occurring in leaf and stem AZs of MirabillisJalapaL., we have expanded our research to tomato, using genomic approaches that include modern techniques for gene discovery and functional gene characterization. In our one-year feasibility study, the US team has established a useful system for VIGS in tomato, using vectors based on the tobacco rattle virus (TRV), a Lcreporter gene for silencing (involved in regulation of anthocyanin biosynthesis), and the gene of interest. In parallel, the Israeli team has used the newly released Affymetrix Tomato GeneChip to measure gene expression in AZ and non-AZ tissues at various time points after flower removal, when increased sensitivity to ethylene is acquired prior to abscission (at 0-8 h), and during pedicelabscission (at 14 h). In addition, gene expression was measured in the pedicel AZ pretreated with the ethylene action inhibitor, 1-methylcyclopropene (1-MCP) before flower removal, to block any direct effects of ethylene. Major conclusions, solutions and achievements: 1) The feasibility study unequivocally established that VIGS is an ideal tool for testing the function of genes with putative roles in abscission; 2) The newly released Affymetrix Tomato GeneChip was found to be an excellent tool to identify AZ genes possibly involved in regulation and execution of abscission. The VIGS-based study allowed us to show that TAPG, a polygalacturonase specifically associated with the tomato AZ, is a key enzyme in the abscission process. Using the newly released Affymetrix Tomato GeneChip we have identified potential abscission regulatory genes as well as new AZ-specific genes, the expression of which was modified after flower removal. These include: members of the Aux/IAAgene family, ethylene signal transduction-related genes, early and late expressed transcription factors, genes which encode post-translational regulators whose expression was modified specifically in the AZ, and many additional novel AZ-specific genes which were previously not associated with abscission. This microarray analysis allowed us to select an initial set of target genes for further functional analysis by VIGS. Implications: Our success in achieving the two objectives of this feasibility study provides us with a solid basis for further research outlined in the original proposal. This will significantly increase the probability of success of a full 3-year project. Additionally, our feasibility study yielded highly innovative results, as they represent the first direct demonstration of the functional involvement of a TAPG in abscission, and the first microarray analysis of the abscission process. Using these approaches we could identify a large number of genes involved in abscission regulation, initiation and execution, and in auxin-ethylene cross-talk, which are of great importance, and could enable their potential functional analysis by VIGS.
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Stern, David B., and Gadi Schuster. Manipulation of Gene Expression in the Chloroplast: Control of mRNA Stability and Transcription Termination. United States Department of Agriculture, 1993. http://dx.doi.org/10.32747/1993.7568750.bard.
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Chloroplasts are the site of photosynthesis and of other essential biosynthetic activities in plant cells. Chloroplasts are semi-autonomous organelles, since they contain their own genomes and protein biosynthetic machinery, but depend on the coordinate expression of nuclear genes to assemble macromolecular complexes. The bioeingineering of plants requires manipulation of chloroplast gene expression, and thus a knowledge of the molecular mechanisms that modulate mRNA and protein production. In this proposal the heterotrophic green alga Chlamydomonas reinhardtii has been used as a model system to understand the control and interrelationships between transcription termination, mRNA 3' end processing and mRNA stability in chloroplasts. Chlamydomonas is a unique and ideal system in which to address these issues, because the chloroplast can be easily manipulated by genetic transformation techniques. This research uncovered new and important information on chloroplast mRNA 3' end formation and mRNA stability. In particular, the 3' untranslated regions of chloroplast mRNAs were shown not to be efficient transcription terminators. The endonucleolytic site in the 3' untranslated region was characterized by site directed mutagensis and the role of several 3' untranslated regions in modulating RNA stability and translation has been studied. This information will allow us to experimentally manipulate the expression of chloroplast genes in vivo by post-transcriptional mechanisms, and should be widely applicable to other higher plant systems.
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Wang, X. F., and M. Schuldiner. Systems biology approaches to dissect virus-host interactions to develop crops with broad-spectrum virus resistance. United States-Israel Binational Agricultural Research and Development Fund, 2020. http://dx.doi.org/10.32747/2020.8134163.bard.
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More than 60% of plant viruses are positive-strand RNA viruses that cause billion-dollar losses annually and pose a major threat to stable agricultural production, including cucumber mosaic virus (CMV) that infects numerous vegetables and ornamental trees. A highly conserved feature among these viruses is that they form viral replication complexes (VRCs) to multiply their genomes by hijacking host proteins and remodeling host intracellular membranes. As a conserved and indispensable process, VRC assembly also represents an excellent target for the development of antiviral strategies that can be used to control a wide-range of viruses. Using CMV and a model virus, brome mosaic virus (BMV), and relying on genomic tools and tailor-made large-scale resources specific for the project, our original objectives were to: 1) Identify host proteins that are required for viral replication complex assembly. 2) Dissect host requirements that determine viral host range. 3) Provide proof-of-concept evidence of a viral control strategy by blocking the viral replication complex-localized phospholipid synthesis. We expect to provide new ways and new concepts to control multiple viruses by targeting a conserved feature among positive-strand RNA viruses based on our results. Our work is going according to the expected timeline and we are progressing well on all aims. For Objective 1, among ~6,000 yeast genes, we have identified 96 hits that were possibly play critical roles in viral replication. These hits are involved in cellular pathways of 1) Phospholipid synthesis; 2) Membrane-shaping; 3) Sterol synthesis and transport; 4) Protein transport; 5) Protein modification, among many others. We are pursuing several genes involved in lipid metabolism and transport because cellular membranes are primarily composed of lipids and lipid compositional changes affect VRC formation and functions. For Objective 2, we have found that CPR5 proteins from monocotyledon plants promoted BMV replication while those from dicotyledon plants inhibited it, providing direct evidence that CPR5 protein determines the host range of BMV. We are currently examining the mechanisms by which dicot CPR5 genes inhibit BMV replication and expressing the dicot CPR5 genes in monocot plants to control BMV infection. For Objective 3, we have demonstrated that substitutions in a host gene involved in lipid synthesis, CHO2, prevented the VRC formation by directing BMV replication protein 1a (BMV 1a), which remodels the nuclear membrane to form VRCs, away from the nuclear membrane, and thus, no VRCs were formed. This has been reported in Journal of Biological Chemistry. Based on the results from Objective 3, we have extended our plan to demonstrate that an amphipathic alpha-helix in BMV 1a is necessary and sufficient to target BMV 1a to the nuclear membrane. We further found that the counterparts of the BMV 1a helix from a group of viruses in the alphavirus-like superfamily, such as CMV, hepatitis E virus, and Rubella virus, are sufficient to target VRCs to the designated membranes, revealing a conserved feature among the superfamily. A joint manuscript describing these exciting results and authored by the two labs will be submitted shortly. We have also successfully set up systems in tomato plants: 1) to efficiently knock down gene expression via virus-induced gene silencing so we could test effects of lacking a host gene(s) on CMV replication; 2) to overexpress any gene transiently from a mild virus (potato virus X) so we could test effects of the overexpressed gene(s) on CMV replication. In summary, we have made promising progress in all three Objectives. We have identified multiple new host proteins that are involved in VRC formation and may serve as good targets to develop antiviral strategies; have confirmed that CPR5 from dicot plants inhibited viral infection and are generating BMV-resistance rice and wheat crops by overexpressing dicot CPR5 genes; have demonstrated to block viral replication by preventing viral replication protein from targeting to the designated organelle membranes for the VRC formation and this concept can be further employed for virus control. We are grateful to BARD funding and are excited to carry on this project in collaboration.