Bibliographies thématiques / Gene functional characterization

Littérature scientifique sur le sujet « Gene functional characterization »

Auteur : Grafiati
Publié le 11 mars 2023

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Articles de revues sur le sujet "Gene functional characterization"

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Jukic, Marin M., Volker M. Lauschke, Takahiro Saito, Masahiro Hiratsuka et Magnus Ingelman-Sundberg. « Functional characterization of CYP2D7 gene variants ». Pharmacogenomics 19, no 12 (août 2018) : 931–36. http://dx.doi.org/10.2217/pgs-2018-0065.

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Ramirez, M. S., T. R. Parenteau, D. Centron et M. E. Tolmasky. « Functional characterization of Tn1331 gene cassettes ». Journal of Antimicrobial Chemotherapy 62, no 4 (27 juin 2008) : 669–73. http://dx.doi.org/10.1093/jac/dkn279.

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KE, Dan-Xia, Kun-Peng PENG, Yan JIA, Shuo ZENG, Ying-Zhi WANG et Jing-Yi ZHANG. « Functional Characterization of Soybean Cystatins Gene GmCYS2 ». Acta Agronomica Sinica 44, no 8 (2018) : 1159. http://dx.doi.org/10.3724/sp.j.1006.2018.01159.

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Thomas, Hugh. « Functional characterization of an IBD risk gene ». Nature Reviews Gastroenterology & ; Hepatology 15, no 4 (21 février 2018) : 191. http://dx.doi.org/10.1038/nrgastro.2018.17.

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Agrawal, N., et M. A. Brown. « Genetic associations and functional characterization of M1 aminopeptidases and immune-mediated diseases ». Genes & ; Immunity 15, no 8 (21 août 2014) : 521–27. http://dx.doi.org/10.1038/gene.2014.46.

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Steffensen, Ane Y., Mette Dandanell, Lars Jønson, Bent Ejlertsen, Anne-Marie Gerdes, Finn C. Nielsen et Thomas vO Hansen. « Functional characterization of BRCA1 gene variants by mini-gene splicing assay ». European Journal of Human Genetics 22, no 12 (26 mars 2014) : 1362–68. http://dx.doi.org/10.1038/ejhg.2014.40.

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Wang, Y., T. M. Hahn, S. Y. Tsai et S. L. Woo. « Functional characterization of a unique liver gene promoter. » Journal of Biological Chemistry 269, no 12 (mars 1994) : 9137–46. http://dx.doi.org/10.1016/s0021-9258(17)37087-4.

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Bocchi, L., T. Fasano, C. Degirolamo, C. Candini, E. Favari, F. Bernini, S. Calandra et S. Bertolini. « PO5-152 FUNCTIONAL CHARACTERIZATION OF ABCA1 GENE MUTANTS ». Atherosclerosis Supplements 8, no 1 (juin 2007) : 55. http://dx.doi.org/10.1016/s1567-5688(07)71162-5.

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Lu, Muxue, Zhigang Wang, Shan Fu, Guangzhe Yang, Mingxing Shi, Youshe Lu, Xiaohu Wang et Jixing Xia. « Functional characterization of the SbNrat1 gene in sorghum ». Plant Science 262 (septembre 2017) : 18–23. http://dx.doi.org/10.1016/j.plantsci.2017.05.010.

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Salman, Alamery, Attia Kotb, Abdelhalim I. Ghazy, Eid I. Ibrahim et Talal K. Al-Ateeq. « Structural and functional characterization of Tomato SUMO1 gene ». Saudi Journal of Biological Sciences 27, no 1 (janvier 2020) : 352–57. http://dx.doi.org/10.1016/j.sjbs.2019.10.004.

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Thèses sur le sujet "Gene functional characterization"

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RIGAMONTI, AURORA. « Functional characterization of SMARCA2 gene ». Doctoral thesis, Università degli Studi di Milano-Bicocca, 2016. http://hdl.handle.net/10281/130275.

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My PhD project is based on the characterization of the SMARCA2 gene. On one hand, I focused my attention on the role of SMARCA2 product, the protein brahma (BRM), in the regulation of alternative pre-mRNA splicing. On the other hand, I investigated the transcriptional regulation of SMARCA2 expression. My first project derived from a comparative evaluation and validation of microarray data from two mitochondrial stress models. The first model represented by an acute mitochondrial stress is constituted by human SH-SY5Y neuroblastoma cells treated with Paraquat (PQ), while the second model is a chronic model constituted from the same cell line stably overexpressing the Superoxide Dismutase 1 carrying the most common mutation found in familiar ALS (SOD1 G93A). The merge of these microarrays data showed that oxidative stress affects the choice of specific alternative last exons (ALEs) increasing the production of transcripts variants terminating at a more proximal ALE. Moreover, oxidative stress induces the transcriptional downregulation of the SMARCA2 gene product BRM, one of the two alternative ATPase subunits of the SWI/SNF complex. I found that in normal condition BRM is enriched on the proximal ALE. In addition, I observed the accumulation of BARD1, a protein that forms a functional heterodimer with BRCA1, which has E3 ubiquitin-ligase activity and interacts with the 50 kDa subunit of CstF inhibiting 3’ end processing. Consistent with these observations, I detected an ubiquitinated pool of CstF50 and showed that ubiquitination is mediated by BARD1/BRCA1. Taken together, these results suggested that the presence of BRM on the proximal exon leads to the BARD1/BRCA1-mediated ubiquitination of CstF50 and the inhibition of 3’ end processing at the proximal poly(A). This in turn allows transcription to proceed to the distal terminal exon. In the same microarray data used as a starting point for my first project we detected a shift in SMARCA2 expression towards shorter mRNA isoforms upon oxidative stress. Thus, my second project dealt with the characterization of these transcripts. Bioinformatic analysis revealed that the shorter mRNA variants are evolutionarily conserved and are most likely generated from an internal promoter. Interestingly, in zebrafish the short isoform is produced as an independent gene on the same chromosome of the long isoform but in its reverse strand. This peculiar genomic organization hints to a potentially relevant function for this alternative isoform of the BRM protein. Bioinformatic analyses revealed that the short isoform encode a protein that lacks the N-terminal, catalytic ATP-ase domain but shares the C-terminal region that contains a Bromodomain, a protein motif that is known to bind to acetylated histones. First, I identified the potential alternative promoter region using bioinformatic tools and cloned this region. Using a luciferase reporter system I demonstrated the existence of the alternative promoter. Next, I cloned the short, most conserved isoform and I tested it for interaction with known partners of the full-length BRM protein by Co-Immunoprecipitation (Co-IP). I discovered that BRM-s interacts with histone H3 but not with core component of the SWI/SNF complex. Considering that short isoform does not display an ATPase domain, the Co-IP suggests a possible “dominant negative” role for this protein. If short isoforms works as negative dominant of Brm, the ratio between long and short expressed proteins could become very important for BRM target genes in development and differentiation. In particular, this alteration of ratio could have a negative effect on the cells since several tumor cell lines show a very low level of the long protein. (i.e. human lung tumor cell lines).
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Lee, Ka Young. « Functional characterization of gene regulation by nhr-49 ». Thesis, University of British Columbia, 2016. http://hdl.handle.net/2429/58365.

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Nuclear hormone receptors (NHRs) are transcription factors that regulate a wide variety of developmental and physiological processes. NHRs are targets of numerous drugs. However, due to limited knowledge on NHR specificity, many such drugs activate multiple biological pathways downstream of NHRs, leading to undesired side effects. To study NHR specificity in vivo, I used the model organism Caenorhabditis elegans. One C. elegans NHR is NHR-49, which regulates various aspects of lipid metabolism. Specifically, it activates genes involved in fatty acid desaturation and fatty acid β-oxidation by binding to a subunit of the Mediator multiprotein complex, MDT-15. Vice versa, NHR-49 represses genes involved in sphingolipid breakdown by heterodimerizing with another C. elegans NHR, NHR-66. Recently, three point mutations in nhr-49 were identified that promote fatty acid desaturation, but whether these alleles act specifically in this pathway or also affect other nhr-49 regulated processes is not clear. To test whether the mutated residues are linked to specific biological functions, I studied how they affect gene expression and protein-protein interactions by real time quantitative PCR and Yeast 2 Hybrid assays. I found that the three point mutations have different effects on nhr-49 dependent metabolic processes. While all three alleles broadly promoted nhr-49 dependent activation, only one allele affected nhr-49 dependent repression. This shows that the mutations and the corresponding amino acid residues have some association with specific nhr-49 dependent biological processes.
Medicine, Faculty of
Medical Genetics, Department of
Graduate
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Pomerleau, Véronique. « Functional characterization of the BHD tumor suppressor gene ». Thesis, McGill University, 2006. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=101734.

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Birt-Hogg-Dube syndrome is a hereditary cancer syndrome that increases the risk of developing kidney cancer. The gene responsible for this disease was recently identified but the protein encoded by this gene has yet to be characterized. In this thesis, we present an initial characterization of the BHD protein. To do so, a polyclonal antibody against the human BHD protein was produced that detects both the exogenous and endogenous protein. Moreover, several BHD putative interacting proteins were identified by multidimensional protein identification technology (MudPIT) analysis following large scale purification of epitope-tagged BHD. The interaction of BHD with one candidate was investigated by co-immunoprecipitation and by in vitro binding assays but could not be validated. In parallel, a Far Western was attempted to discover BHD putative interacting proteins but with no success. Furthermore, the intracellular localization of BHD revealed to be mostly cytoplasmic as shown by differential centrifugation and by indirect immunofluorescence. A BHDwt-GFP construct was also engineered to perform further studies on BHD subcellular localization. Finally, the effect of BHD disease-associated mutations on protein expression was investigated by Western blotting.
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Dennis, Megan Young. « Functional characterization of the dyslexia candidate gene KIAA0319 ». Thesis, University of Oxford, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.504385.

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Le, Hang Thi Thu. « Functional characterization of IGF2BP2, a diabetes-susceptibility gene ». Thesis, University of Cambridge, 2011. https://www.repository.cam.ac.uk/handle/1810/283871.

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Wahab, S. M. Riajul. « Molecular and functional characterization of GAEC1 gene in human colorectal cancer ». Thesis, Griffith University, 2018. http://hdl.handle.net/10072/377621.

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Background: Colorectal cancer is now one of the most common causes of death in Australia, with an estimated 1486 new cases in the country in 2010, accounting 12.7% of all cancer deaths (ACIM, 2014). In addition to its significance in Australia, it is one of the most common global health concerns. At present colorectal cancer is the third most common cancer worldwide, which cost more than 600,000 lives every year. Most of the colorectal cancer is diagnosed at a late stage but if it is diagnosed at an early stage, the five-year survival rate exceeds in 90% cases. This is the reason there is a need to find out biomarker for early detection and the exact underlying cause for designing a better treatment for colorectal cancer. GAEC1 (Gene amplified in esophageal cancer 1) showed a series of amplifications and deletions in oesophageal cancer. The gene is located at 7q22.1. GAEC1 has tumorigenic potential approximately equal to the Ras gene family and overexpression of this gene played a pivotal role in the cancer transformation of oesophageal squamous cell carcinoma. GAEC1 has higher amplification in colorectal adenocarcinoma tissues when compared to non-cancer colorectal tissues. In this study, we focused on finding out the oncogenic properties of GAEC1, correlation with clinical and pathological features and its underlying mechanism in colorectal cancer initiation and progression. Materials and method: Human colon cancer cell lines (SW480, SW48, HCT116 cells) and non-neoplastic colonic epithelium cell (FHC cells) were purchased from American Type Culture Collection (ATCC). SW480, SW48 and HCT116 cell lines were maintained in Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum at 37 ℃ in 5% CO2. FHC cells were maintained in DMEM: F-12 (1:1) with 10% fetal bovine serum with containing an extra 10 mM N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) (Thermo Fisher Scientific) (for a final concentration of 25 mM), 10 ng/ml cholera toxin, 0.005 mg/ml insulin, 0.005 mg/ml transferrin, 100 ng/ml hydrocortisone. Fresh frozen human colorectal cancer tissues and adjacent non-cancer tissues were collected with no selection bias. Expression levels of mRNA and protein were measured by real-time PCR and western blot analysis respectively. Immunocytochemistry, immunohistochemistry and immunofluorescence assay were used to identify the localization of GAEC1 protein in colon cancer cells and colon cancer tissues. Flow cytometry was used for the detection of apoptotic cells and cell cycle alteration. Co-immunoprecipitation followed by mass spectrometry analysis was used to identify the protein-protein interaction. Severe combined immunodeficiency (SCID) mice were used for tumour xenograft experiment. Results: We found differential expression of GAEC1 protein and mRNA in different pathological stages of colon cancer cells (SW480-Stage II, SW48-Stage III and HCT116-Stage IV) when compared to non-neoplastic colon cells (FHC cells). GAEC1 protein was predominantly expressed in the cytoplasm of colon cancer cells (SW480, SW48, and HCT116) and the nucleus of non-neoplastic colon epithelial cells (FHC). The transient knockdown of GAEC1 using siRNA induced apoptosis in SW480 and SW48 cells, which was associated with G2/M phase arrest and decreased expression of Bcl-2 and K-ras proteins and increased expression of p53. In addition, down-regulation of GAEC1 significantly inhibited cell proliferation, reduced migration capacity and decreased clonogenic potentiality of colon cancer cells (SW480 and SW48 cells). Furthermore, a xenotransplantation model showed that stable knockdown of GAEC1 using shRNA constructs in colon cancer cells entirely suppressed xenograft tumour growth in mice. Approximately 52.5% of patients with colorectal cancers showed high expression of GAEC1 mRNA whereas 47.5% exhibited low expression compared to their matched non-neoplastic tissues. Similarly, ~ 66% (53/80) of colorectal cancer tissues showed high GAEC1 protein expression (positive staining), while the remaining colorectal cancer cases were noted with no GAEC1 protein (negative) expression. GAEC1 protein was predominantly located in the cytoplasm and showed low to no expression in normal colon tissues. High expression of GAEC1 mRNA was predominantly seen among patients below 60 years compared to those patients over 60 years of age (78%, versus 44%, p=0.008). Patients with synchronous colorectal adenocarcinomas mostly exhibited with low expression of GAEC1 mRNA. On the other hand, compared to poorly differentiated colorectal carcinomas (grade III), patients with well and moderately differentiated colorectal carcinomas (grade I+II) colorectal cancers showed a high expression of GAEC1 mRNA. Similarly, high GAEC1 mRNA expression was frequently noted among patients presented without any pre-neoplastic adenomas in their colorectal cancer tissues compared to patients with an adenoma in their colorectal cancer tissues. By co-immunoprecipitation followed by mass spectrometry analysis 31 interacting protein was identified. The interaction between GAEC1 and four proteins (HIGD1A, Rhotekin, Granulin and eIF3J) was further confirmed. Western blot analysis detected reduced expression of these proteins following stable knockdown of GAEC1 in colon cancer cells. GEAC1 endogenously interacts with p53 in SW480 and SW48 colon cancer cells. In this study, we have noted that overexpression of GAEC1 increased cell proliferation, migration, and reduced apoptosis in colon cancer cells. Also, these cells showed cell cycle arrest at the synthetic phase, activation of Bcl-2, K-ras, pAKT proteins as well as inhibition of p53, PUMA, p21 and BAX proteins. Furthermore, silencing of GAEC1 reduces the nuclear import of MDM2 and increase the expression of p53 in the nucleus suggesting that GAEC1 expression is essential for interaction of p53-MDM2 and nuclear translocation of MDM2 in colon cancer cells. Conclusion: In summary, the expression analysis, in vitro and in vivo data indicated that GAEC1 is differentially expressed in cancer cells and act as an oncogene in colon cancer progression. The high expression of GAEC1 mRNA/protein, as well as its correlation with multiple clinical and pathological characteristics in patients with colorectal carcinoma, strongly, suggests that GAEC1 is a key regulator in the initiation of colorectal carcinogenesis. In addition, the protein-protein interaction with a number of proteins and the effect of GAEC1 modulation on the expression of interacting proteins indicates the potential role of GAEC1 in the signalling pathway of colon cancer pathogenesis.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Medicine
Griffith Health
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Zhang, Xue-Cheng. « Functional characterization of the Arabidopsis disease resistance gene RPS4 ». Diss., Columbia, Mo. : University of Missouri-Columbia, 2005. http://hdl.handle.net/10355/5826.

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Thesis (Ph.D.)--University of Missouri-Columbia, 2005.
The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file viewed on (November 27, 2006) Vita. Includes bibliographical references.
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Salehin, Mohammad. « Molecular and Functional Characterization of Medicago Truncatula Npf17 Gene ». Thesis, University of North Texas, 2013. https://digital.library.unt.edu/ark:/67531/metadc407747/.

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Legumes are unique among plants for their ability to fix atmospheric nitrogen with the help of soil bacteria rhizobia. Medicago truncatula is used as a model legume to study different aspects of symbiotic nitrogen fixation. M. truncatula, in association with its symbiotic partner Sinorhizobium meliloti, fix atmospheric nitrogen into ammonia, which the plant uses for amino acid biosynthesis and the bacteria get reduced photosynthate in return. M. truncatula NPF1.7 previously called MtNIP/LATD is required for symbiotic nitrogen fixing root nodule development and for normal root architecture. Mutations in MtNPF1.7 have defects in these processes. MtNPF1.7 encodes a member of the NPF family of transporters. Experimental results showing that MtNPF1.7 functioning as a high-affinity nitrate transporter are its expression restoring chlorate susceptibility to the Arabidopsis chl1-5 mutant and high nitrate transport in Xenopus laevis oocyte system. However, the weakest Mtnip-3 mutant allele also displays high-affinity nitrate transport in X. laevis oocytes and chlorate susceptibility to the Atchl1-5 mutant, suggesting that MtNPF1.7 might have another biochemical function. Experimental evidence shows that MtNPF1.7 also functions in hormone signaling. Constitutive expression of MtNPF1.7 in several species including M. truncatula results in plants with a robust growth phenotype. Using a synthetic auxin reporter, the presence of higher auxin in both the Mtnip-1 mutant and in M. truncatula plants constitutively expressing MtNPF1.7 was observed. Previous experiments showed MtNPF1.7 expression is hormone regulated and the MtNPF1.7 promoter is active in root and nodule meristems and in the vasculature. Two potential binding sites for an auxin response factors (ARFs) were found in the MtNPF1.7 promoter. Chromatin immunoprecipitation-qRT-PCR confirmed MtARF1 binding these sites. Mutating the MtARF1 binding sites increases MtNPF1.7 expression, suggesting a mechanism for auxin repression of MtNPF1.7. Consistent with these results, constitutive expression of an ARF in wild-type plants partially phenocopies Mtnip-1 mutants’ phenotypes.
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Crimmins, Stephen Lewis. « Characterization and functional analysis of Usp14 ». Thesis, Birmingham, Ala. : University of Alabama at Birmingham, 2007. https://www.mhsl.uab.edu/dt/2007p/crimmins.pdf.

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Yan, Kaiping. « Characterization and functional analyses of the transcriptional cofactor TIF1γ gene ». Université Louis Pasteur (Strasbourg) (1971-2008), 2003. http://www.theses.fr/2003STR13144.

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Livres sur le sujet "Gene functional characterization"

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Orozco, Ian Jay. Functional Characterization of Hippocampal Synapses in a Mouse Mutant of the Dystrobrevin Binding Protein 1 (DTNBP 1) Gene. [New York, N.Y.?] : [publisher not identified], 2011.

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Roepke, Jonathon. Localization and functional characterization of iridoid biosynthetic genes in Catharanthus roseus. St. Catharines, Ont : Brock University, Centre for Biotechnology, 2008.

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A, Nicolini Claudio, North Atlantic Treaty Organization. Scientific Affairs Division. et NATO Advanced Study Institute on Genome Structure and Function (1996 : Marciana Marina, Italy), dir. Genome structure and function : From chromosomes characterization to genes technology. Dordrecht : Kluwer, 1997.

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Huang, Weei-Yuarn. Nucleosomal structure and functions : Characterization of the hamster cardiac myosin heavy chain genes DNase I hypersensitive sites. Ottawa : National Library of Canada = Bibliothèque nationale du Canada, 1997.

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He, Peng, Nicolas Durand et Shuang-Lin Dong, dir. Insect Olfactory Proteins (From Gene Identification to Functional Characterization). Frontiers Media SA, 2020. http://dx.doi.org/10.3389/978-2-88963-266-4.

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He, Peng, Xiaojiao Guo, Yang Liu, Wei Xu, Jin Zhang, Ya-Nan Zhang et J. Joe Hull, dir. Insect Olfactory Proteins (From Gene Identification to Functional Characterization), Volume II. Frontiers Media SA, 2022. http://dx.doi.org/10.3389/978-2-88974-749-8.

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To, Minh Dong. Genetic and functional characterization of human presenilin-2 gene involvement in breast and colon cancer. 2003.

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Turley, Stefanie A. Functional characterization of PRDM12, a gene recurrently deleted during t(9;22) rearrangements in chronic myeloid leukemia patients. 2006.

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Garrad, Richard C. The molecular and functional analysis of the LYS1 gene of Candida albicans and characterization of lysine auxotrophs of Candida spp. 1993.

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Koller, Hannelore T. Multiple functional in vivo spacer rRNA gene promoters during early development and characterization of the intergenic spacer in the crustacean Artemia. 1987.

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Chapitres de livres sur le sujet "Gene functional characterization"

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Reid, Michael, Jen-Chih Chen et Cai-Zhong Jiang. « Virus-Induced Gene Silencing for Functional Characterization of Genes in Petunia ». Dans Petunia, 381–94. New York, NY : Springer New York, 2009. http://dx.doi.org/10.1007/978-0-387-84796-2_18.

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Guo, Hui-jun, Yong-dun Xie, Lin-shu Zhao, Hong-chun Xiong, Jia-yu Gu, Shi-rong Zhao et Lu-xiang Liu. « Progress of mutant resource development and tilling on starch biosynthesis in wheat. » Dans Mutation breeding, genetic diversity and crop adaptation to climate change, 280–84. Wallingford : CABI, 2021. http://dx.doi.org/10.1079/9781789249095.0028.

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Abstract Induced mutations have been widely utilized for the development of plant mutant germplasm and varieties since 1927 and have contributed to genetic diversity enhancement and food security in the world. Mutant resources are essential for gene identification and functional characterization by forward and reverse genetic strategies. The publishing of annotated wheat reference genomes is greatly promoting the progress of wheat functional genomic research. Mutant resources of a broad spectrum and diversified wild- types will be the prerequisites in this process, in part due to the polyploid nature of wheat. This review describes the progress of mutant resource development derived from the winter wheat cultivar 'Jing411'. The segregating M2 population has been used for mining functional mutant alleles of key genes involved in starch biosynthesis and could be further used for allele mining of any other target genes. The morphological mutant resources developed from various mutagens have been, and are going to be, used to develop genetic populations for gene mapping and the genetic analysis of biological functions.
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Pont, Suzanne, Anne Regnier-Vigouroux, Philippe Naquet et Michel Pierres. « Functional Characterization of Several Steric Domains of the Murine Thy-1 and LFA-1 Molecules ». Dans Regulation of Immune Gene Expression, 111–17. Totowa, NJ : Humana Press, 1986. http://dx.doi.org/10.1007/978-1-4612-5014-2_10.

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Saha, Mousumi, Agniswar Sarkar et Bidyut Bandyopadhyay. « Phylogenetic Characterization of Nitrifying Bacteria Isolated from East Kolkata Wetland ». Dans Proceedings of the Conference BioSangam 2022 : Emerging Trends in Biotechnology (BIOSANGAM 2022), 114–22. Dordrecht : Atlantis Press International BV, 2022. http://dx.doi.org/10.2991/978-94-6463-020-6_12.

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AbstractEast Kolkata Wetland (EKW) is an “International Ramsar Site”, famous for broad biodiversity and insightful use of sewage for aquaculture. Native nitrifying bacteria of EKW play a significant role in maintaining water quality and controlling environmental pollution by converting ammonia into nitrate in wastewater. Therefore, the characterization of nitrifying bacteria is important in EKW. Thus, the main focus of this research was to identify and characterize the nitrifying bacteria, investigating their phylogeny and diversity in EKW. 16S rRNA and functional genes analysis may help in the proper evaluation of composition and distribution of nitrifying bacteria in some water bodies in EKW, which has not yet been explored. Molecular and phylogenetic characterization was targeted and achieved through 16S rRNA and functional gene analysis, followed by computational estimation. Resulted sequences were analysed to gain insight into the knowledge for global and local taxonomic orientation. Hence, a model can be created for characterizing the dynamics of nitrifying bacteria in wastewater treatment and sustainable aquaculture in different water bodies of EKW. Graphical Abstract
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Jung, Ki-Hong, et Gynheung An. « Functional Characterization of Rice Genes Using a Gene-Indexed T-DNA Insertional Mutant Population ». Dans Methods in Molecular Biology, 57–67. Totowa, NJ : Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-194-3_5.

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Gunupuru, Lokanadha R., Alexandre Perochon, Shahin S. Ali, Steven R. Scofield et Fiona M. Doohan. « Virus-Induced Gene Silencing (VIGS) for Functional Characterization of Disease Resistance Genes in Barley Seedlings ». Dans Methods in Molecular Biology, 95–114. New York, NY : Springer New York, 2018. http://dx.doi.org/10.1007/978-1-4939-8944-7_7.

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Wullems, George J., Floyd R. A. Wittink, Peter de Groot, Jan Derksen et Jan A. M. Schrauwen. « Characterization of a pollen specific gene and its functional analysis in transgenic plants ». Dans Plant Biotechnology and In Vitro Biology in the 21st Century, 235–39. Dordrecht : Springer Netherlands, 1999. http://dx.doi.org/10.1007/978-94-011-4661-6_55.

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Port, Fillip, et Michael Boutros. « Tissue-Specific CRISPR-Cas9 Screening in Drosophila ». Dans Methods in Molecular Biology, 157–76. New York, NY : Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2541-5_7.

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AbstractOver the last century research in Drosophila has resulted in many fundamental contributions to our understanding of the biology of multicellular organisms. Many of these breakthroughs have been based on the identification of novel gene functions in large-scale genetic screens. However, conventional forward-genetic screens have been limited by the random nature of mutagenesis and difficulties in mapping causal mutations, while reverse-genetic RNAi screens suffer from incomplete knockdown of gene expression. Recently developed large-scale CRISPR-Cas9 libraries promise to address these limitations by allowing the induction of targeted mutations in genes with spatial and temporal control. Here, we provide a guide for tissue-specific CRISPR screening in Drosophila, including the characterization of Gal4 UAS-Cas9 lines, selection of sgRNA libraries, and various quality control measures. We also discuss confounding factors that can give rise to false-positive and false-negative results in such experiments and suggest strategies on how to detect and avoid them. Conditional CRISPR screening represents an exciting new approach for functional genomics in vivo and is set to further expand our knowledge of the molecular underpinning of development, homeostasis, and disease.
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Ball, Edward J., Kay Black, Marie Hoover, J. Donald Capra et Peter Stastny. « Functional Characterization of T-cell Clones Using Related T-Cell Receptor V Gene Segments ». Dans Immunobiology of HLA, 550–51. Berlin, Heidelberg : Springer Berlin Heidelberg, 1989. http://dx.doi.org/10.1007/978-3-662-39946-0_237.

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Decker, H. J. H., S. Klauck, D. I. Smith, C. Huber et B. Seizinger. « Characterization and Functional Significance of the von Hippel-Lindau Gene in Renal Cell Carcinomas ». Dans Contemporary Research on Renal Cell Carcinoma, 97–103. Berlin, Heidelberg : Springer Berlin Heidelberg, 1994. http://dx.doi.org/10.1007/978-3-642-78609-9_12.

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Actes de conférences sur le sujet "Gene functional characterization"

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Kuuselo, Riina, Kimmo Savinainen et Anne Kallioniemi. « Abstract 248 : Functional characterization of MED29 gene in pancreatic cancer ». Dans Proceedings : AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010 ; Washington, DC. American Association for Cancer Research, 2010. http://dx.doi.org/10.1158/1538-7445.am10-248.

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Yalçın Çapan, Özlem. « Functional Characterization of p.T10M and p.S345Y mutations in HNF1A Gene in MODY patients ». Dans 15th International Congress of Histochemistry and Cytochemistry. Istanbul : LookUs Scientific, 2017. http://dx.doi.org/10.5505/2017ichc.op-25.

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Bhattacharjee, Vikram, et Timothy Yen. « Abstract A10 : Functional characterization of gene sensitizers of pancreatic cancer cells to gemcitabine. » Dans Abstracts : AACR Special Conference on Pancreatic Cancer : Progress and Challenges ; June 18-21, 2012 ; Lake Tahoe, NV. American Association for Cancer Research, 2012. http://dx.doi.org/10.1158/1538-7445.panca2012-a10.

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Wang, Yanjun, Qian Yang et Jinzhu Song. « Expression and Functional Characterization of Chitinase Gene, CHI58 Related to Biocontrol in Chaetomium cupreum ». Dans 2009 3rd International Conference on Bioinformatics and Biomedical Engineering (iCBBE 2009). IEEE, 2009. http://dx.doi.org/10.1109/icbbe.2009.5162831.

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Conboy, Caitlin B., Julia M. Hatler, Pat M. Scott, Aaron L. Sarver, Kevin A. Silverstein, Somasekar Seshagiri, Matthew Adams et al. « Abstract 108 : Functional characterization of WAC, a candidate tumor suppressor gene in colorectal cancer ». Dans Proceedings : AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012 ; Chicago, IL. American Association for Cancer Research, 2012. http://dx.doi.org/10.1158/1538-7445.am2012-108.

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Koide, T. « CHARACTERIZATION OF THE GENE FOR HUMAN HISTIDINE-RICH GLYCOPROTEIN ». Dans XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643599.

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Human histidine-rich glycoprotein (HRG) is a single-chain glycoprotein in plasma which is considered to modulate a coagulation and fibrinolysis system with the ability to bind to heparin, plasminogen, fibrinogen, thrombospondin, etc. Recently we have elucidated the primary structure of HRG by determining the nucleotide sequence of its cDNA, and showed that HRG is composed of several different types of internal repeats, each one of which shows considerable homology with the functional and/or structural domains of other proteins including high molecular weight kininogen, antithrombin III, cystatins, and proline-rich protein and peptide. Thus, the multifunctional property of HRG was suggested to be due to its multi-domain structure. In the present studies, a human genomic DNA library, cloned in the bacteriophage vector Charon 4A, was screened for HRG gene using a full-length cDNA coding for human IMI as a probe. A total of 7 clones were isolated from 6 × 105 phage and each was plaque purified. The entire HRG gene is represented in 3 genomic inserts with overlapping sequences that carry human DNA spanning 30 kb. Overlapping gene fragments were subcloned into pUC9 and characterized by Southern blot hybridization using 5’ and 3’ end probes isolated from human HRG cDNA and by DNA sequencing. These studies have shown that the gene for human HRG spans about 9 kb and consists of at least 5 exons and 4 introns. The putative histidine-rich region consisted of 12 tandemly repeated sequences of a 5 amino acid segment and 2 proline-rich regions contiguous to it are likely to be involved within one exon.
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Piscuoglio, Salvatore, Michal Kovac, Shivendra Kishore, Valentina Mele, Mihaela Zavolan, Hansjakob Müller, Luigi M. Terracciano et Karl Heinimann. « Abstract 3802 : EWSR1 : Identification and functional characterization of a novel target gene locus in Lynch syndrome ». Dans Proceedings : AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011 ; Orlando, FL. American Association for Cancer Research, 2011. http://dx.doi.org/10.1158/1538-7445.am2011-3802.

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Leong, Merrin Man Long, Arthur Kwok Leung Cheung, Tommy Chin Tung Kwok et Maria Li Lung. « Abstract 5521 : Functional characterization of a candidate tumor suppressor gene,Mirror image polydactyly 1, in nasopharyngeal carcinoma ». Dans Proceedings : AACR Annual Meeting 2018 ; April 14-18, 2018 ; Chicago, IL. American Association for Cancer Research, 2018. http://dx.doi.org/10.1158/1538-7445.am2018-5521.

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Singh, Kamaleshwar P., Shubhashish Sarkar et Deodutta Roy. « Abstract 5050 : Functional characterization of a newly identified growth regulatory gene SKCG-1 in human kidney cancer ». Dans Proceedings : AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010 ; Washington, DC. American Association for Cancer Research, 2010. http://dx.doi.org/10.1158/1538-7445.am10-5050.

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Kwabi-Addo, Bernard, Songping Wang, Paulett Furbert-Harris, Srinivasan Yegnasubramanian et Joseph Devaney. « Abstract 1974 : Functional characterization of Basonuclin 1 (BNC1) : a novel tumor suppressor gene commonly downregulated in human prostate cancer. » Dans Proceedings : AACR 104th Annual Meeting 2013 ; Apr 6-10, 2013 ; Washington, DC. American Association for Cancer Research, 2013. http://dx.doi.org/10.1158/1538-7445.am2013-1974.

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Rapports d'organisations sur le sujet "Gene functional characterization"

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Wang, Ruoxiang. Functional Characterization of a Novel Prostate-Specific Gene PrLZ in Prostate Cancer. Fort Belvoir, VA : Defense Technical Information Center, janvier 2006. http://dx.doi.org/10.21236/ada609546.

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Busov, Victor. Functional Gene Discovery and Characterization of Genes and Alleles Affecting Wood Biomass Yield and Quality in Populus. Office of Scientific and Technical Information (OSTI), février 2017. http://dx.doi.org/10.2172/1343386.

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Dougherty, Shona. Isolation and Functional Characterization of Prostate Tumor-Specific Hypoxia-Inducible Promoter/Enhancer Elements for Use in Gene Therapy. Fort Belvoir, VA : Defense Technical Information Center, juin 2001. http://dx.doi.org/10.21236/ada413596.

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Li, Li, Joseph Burger, Nurit Katzir, Yaakov Tadmor, Ari Schaffer et Zhangjun Fei. Characterization of the Or regulatory network in melon for carotenoid biofortification in food crops. United States Department of Agriculture, avril 2015. http://dx.doi.org/10.32747/2015.7594408.bard.

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The general goals of the BARD research grant US-4423-11 are to understand how Or regulates carotenoid accumulation and to reveal novel strategies for breeding agricultural crops with enhanced β-carotene level. The original objectives are: 1) to identify the genes and proteins in the Or regulatory network in melon; 2) to genetically and molecularly characterize the candidate genes; and 3) to define genetic and functional allelic variation of these genes in a representative germplasm collection of the C. melo species. Or was found by the US group to causes provitamin A accumulation in chromoplasts in cauliflower. Preliminary genetic study from the Israeli group revealed that the melon Or gene (CmOr) completely co-segregated with fruit flesh color in a segregating mapping population and in a wide melon germplasm collection, which set the stage for the funded research. Major conclusions and achievements include: 1). CmOris proved to be the gene that controls melon fruit flesh color and represents the previously described gflocus in melon. 2). Genetic and molecular analyses of CmOridentify and confirm a single SNP that is responsible for the orange and non-orange phenotypes in melon fruit. 3). Alteration of the evolutionarily conserved arginine in an OR protein to both histidine or alanine greatly enhances its ability to promote carotenoid accumulation. 4). OR promotes massive carotenoid accumulation due to its dual functions in regulating both chromoplast biogenesis and carotenoid biosynthesis. 5). A bulk segregant transcriptome (BSRseq) analysis identifies a list of genes associated with the CmOrregulatory network. 6). BSRseq is proved to be an effective approach for gene discovery. 7). Screening of an EMS mutation library identifies a low β mutant, which contains low level of carotenoids due to a mutation in CmOrto produce a truncated form of OR protein. 8). low β exhibits lower germination rate and slow growth under salt stress condition. 9). Postharvest storage of fruit enhances carotenoid accumulation, which is associated with chromoplast development. Our research uncovers the molecular mechanisms underlying the Or-regulated high level of carotenoid accumulation via regulating carotenoidbiosynthetic capacity and storage sink strength. The findings provide mechanistic insights into how carotenoid accumulation is controlled in plants. Our research also provides general and reliable molecular markers for melon-breeding programs to select orange varieties, and offers effective genetic tools for pro-vitamin A enrichment in other important crops via the rapidly developed genome editing technology. The newly discovered low β mutant could lead to a better understanding of the Or gene function and its association with stress response, which may explain the high conservation of the Or gene among various plant species.
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Litovchick, Larisa. Functional Characterization of the Protein Product of the Prostate Carcinoma Gene Fusion TMPRSS2:ERG Using the Proteomic and Microarray Analyses. Fort Belvoir, VA : Defense Technical Information Center, juillet 2009. http://dx.doi.org/10.21236/ada509751.

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Meir, Shimon, Michael Reid, Cai-Zhong Jiang, Amnon Lers et 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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Lers, Amnon, E. Lomaniec, S. Burd, A. Khalchitski, L. Canetti et Pamela J. Green. Analysis of Senescence Inducible Ribonuclease in Tomato : Gene Regulation and Function. United States Department of Agriculture, février 2000. http://dx.doi.org/10.32747/2000.7570563.bard.

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Natural leaf senescence has a negative influence on yield. Postharvest induced senescence contributes to the losses of quality in flowers, foliage, and vegetables. Strategies designed to control the senescence process in crop plants could therefore have great applied significance. Senescence is regulated by differential gene expression yet, functional characterization of the genes specifically induced and study of their expression control, is still in its infancy. Study of senescence-specific genes is required to allow identification of regulatory elements participating in senescence-induced expression and thus provide insights into the genetic regulation of senescence. A main feature of senescence is the hydrolysis of macromolecules by hydrolases of various types such as RNases and proteases. This study was aimed a analysis of senescence-inducible RNases in tomato with the following objectives: Isolation of senescence-inducible RNase cDNA clones; Expression analyses of RNase genes during senescence; Identification of sequences required for senescence-induced gene expression; Functional analyses of senescence-inducible RNases. We narrowed our aims somewhat to focus on the first three objectives because the budget we were awarded was reduced from that requested. We have expanded our research for identification senescence-related RNase/nuclease activities as we thought it will direct us to new RNase/nuclease genes. We have also carried out research in Arabidopsis and parsley, which enabled us to draw mire general conclusions. We completed the first and second objectives and have made considerable progress on the remaining two. We have defined growth conditions suitable for this research and defined the physiological and biochemical parameters characteristic to the advance of leaf senescence. In tomato and arabidopsis we have focused on natural leaf senescence. Parsley was used mainly for study of postharvest senescence in detached leaves. We have identified a 41-kD a tomato nuclease, LeNUCI, specifically induced during senescence which can degrade both RNA and DNA. This activity could be induced by ethylene in young leaves and was subjected to detailed analysis, which enabled its classification as Nuclease I enzyme. LeNUCI may be involved in nucleic acid metabolism during tomato leaf senescence. In parsley senescing leaves we identified 2 main senescence-related nuclease activities of 41 and 39-kDa. These activities were induced in both naturally or artificially senescing leaves, could degrade both DNA and RNA and were very similar in their characteristics to the LeNUCI. Two senescence-induced RNase cDNAs were cloned from tomato. One RNase cDNA was identical to the tomato LX RNase while the second corresponded to the LE RNase. Both were demonstrated before to be induced following phosphate starvation of tomato cell culture but nothing was known about their expression or function in plants. LX gene expression was much more senescence specific and ethylene could activate it in detached young leaves. LE gene expression, which could be transiently induced by wounding, appeared to be activated by abscisic acid. We suggest that the LX RNase has a role in RNA catabolism in the final stage of senescence, and LE may be a defense-related protein. Transgenic plants were generated for altering LX gene expression. No major visible alterations in the phenotype were observed so far. Detailed analysis of senescence in these plants is performed currently. The LX promoter was cloned and its analysis is performed currently for identification of senescence-specific regulatory elements. In Arabidopsis we have identified and characterized a senescence-associated nuclease 1 gene, BFN1, which is highly expressed during leaf and stem senescence. BFN1, is the first example of a senescence- associated gene encoding a nuclease I enzyme as well as the first nuclease I cloned and characterized from Arabidopsis. Our progress should provide excellent tools for the continued analysis of regulation and function of senescence-inducible ribonucleases and nucleases in plants. The cloned genes can be used in reverse genetic approaches, already initiated, which can yield a more direct evidence for the function of these enzymes. Another contribution of this research will be in respect to the molecular mechanism, which controls senescence. We had already initiated in this project and will continue to identify and characterize regulatory elements involved in senescence-specific expression of the genes isolated in this work.
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Dubcovsky, Jorge, Tzion Fahima, Ann Blechl et Phillip San Miguel. Validation of a candidate gene for increased grain protein content in wheat. United States Department of Agriculture, janvier 2007. http://dx.doi.org/10.32747/2007.7695857.bard.

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High Grain Protein Content (GPC) of wheat is important for improved nutritional value and industrial quality. However, selection for this trait is limited by our poor understanding of the genes involved in the accumulation of protein in the grain. A gene with a large effect on GPC was detected on the short arm of chromosome 6B in a Triticum turgidum ssp. dicoccoides accession from Israel (DIC, hereafter). During the previous BARD project we constructed a half-million clones Bacterial Artificial Chromosome (BAC) library of tetraploid wheat including the high GPC allele from DIC and mapped the GPC-B1 locus within a 0.3-cM interval. Our long-term goal is to provide a better understanding of the genes controlling grain protein content in wheat. The specific objectives of the current project were to: (1) complete the positional cloning of the GPC-B1 candidate gene; (2) characterize the allelic variation and (3) expression profile of the candidate gene; and (4) validate this gene by using a transgenic RNAi approach to reduce the GPC transcript levels. To achieve these goals we constructed a 245-kb physical map of the GPC-B1 region. Tetraploid and hexaploid wheat lines carrying this 245-kb DIC segment showed delayed senescence and increased GPC and grain micronutrients. The complete sequencing of this region revealed five genes. A high-resolution genetic map, based on approximately 9,000 gametes and new molecular markers enabled us to delimit the GPC-B1 locus to a 7.4-kb region. Complete linkage of the 7.4-kb region with earlier senescence and increase in GPC, Zn, and Fe concentrations in the grain suggested that GPC-B1 is a single gene with multiple pleiotropic effects. The annotation of this 7.4-kb region identified a single gene, encoding a NAC transcription factor, designated as NAM-B1. Allelic variation studies demonstrated that the ancestral wild wheat allele encodes a functional NAC transcription factor whereas modern wheat varieties carry a non-functional NAM-B1 allele. Quantitative PCR showed that transcript levels for the multiple NAMhomologues were low in flag leaves prior to anthesis, after which their levels increased significantly towards grain maturity. Reduction in RNA levels of the multiple NAMhomologues by RNA interference delayed senescence by over three weeks and reduced wheat grain protein, Zn, and Fe content by over 30%. In the transgenic RNAi plants, residual N, Zn and Fe in the dry leaves was significantly higher than in the control plants, confirming a more efficient nutrient remobilization in the presence of higher levels of GPC. The multiple pleiotropic effects of NAM genes suggest a central role for these genes as transcriptional regulators of multiple processes during leaf senescence, including nutrient remobilization to the developing grain. The cloning of GPC-B1 provides a direct link between the regulation of senescence and nutrient remobilization and an entry point to characterize the genes regulating these two processes. This may contribute to their more efficient manipulation in crops and translate into food with enhanced nutritional value. The characterization of the GPC-B1 gene will have a significant impact on wheat production in many regions of the world and will open the door for the identification of additional genes involved in the accumulation of protein in the grain.
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Dubcovsky, Jorge, Tzion Fahima et Ann Blechl. Molecular characterization and deployment of the high-temperature adult plant stripe rust resistance gene Yr36 from wheat. United States Department of Agriculture, novembre 2013. http://dx.doi.org/10.32747/2013.7699860.bard.

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Stripe rust, caused by Puccinia striiformis f. sp. tritici is one of the most destructive fungal diseases of wheat. Virulent races that appeared within the last decade caused drastic cuts in yields. The incorporation of genetic resistance against this pathogen is the most cost-effective and environmentally friendly solution to this problem. However, race specific seedling resistance genes provide only a temporary solution because fungal populations rapidly evolve to overcome this type of resistance. In contrast, high temperature adult plant (HTAP) resistance genes provide a broad spectrum resistance that is partial and more durable. The cloning of the first wheat HTAP stripe rust resistance gene Yr36 (Science 2009, 323:1357), funded by our previous (2007-2010) BARD grant, provided us for the first time with an entry point for understanding the mechanism of broad spectrum resistance. Two paralogous copies of this gene are tightly linked at the Yr36 locus (WKS1 and WKS2). The main objectives of the current study were to characterize the Yr36 (WKS) resistance mechanism and to identify and characterize alternative WKSgenes in wheat and wild relatives. We report here that the protein coded by Yr36, designated WKS1, that has a novel architecture with a functional kinase and a lipid binding START domain, is localized to chloroplast. Our results suggest that the presence of the START domain may affect the kinase activity. We have found that the WKS1 was over-expressed on leaf necrosis in wheat transgenic plants. When the isolated WKS1.1 splice variant transcript was transformed into susceptible wheat it conferred resistance to stripe rust, but the truncated variant WKS1.2 did not confer resistance. WKS1.1 and WKS1.2 showed different lipid binding profiling. WKS1.1 enters the chloroplast membrane, while WKS1.2 is only attached outside of the chloroplast membrane. The ascorbate peroxidase (APX) activity of the recombinant protein of TmtAPXwas found to be reduced by WKS1.1 protein in vitro. The WKS1.1 mature protein in the chloroplast is able to phosphorylate TmtAPXprotein in vivo. WKS1.1 induced cell death by suppressing APX activity and reducing the ability of the cell to detoxify reactive oxygen. The decrease of APX activity reduces the ability of the plant to detoxify the reactive H2O2 and is the possible mechanism underlying the accelerated cell death observed in the transgenic plants overexpressing WKS1.1 and in the regions surrounding a stripe rust infection in the wheat plants carrying the natural WKS1.1 gene. WKS2 is a nonfunctional paralog of WKS1 in wild emmer wheat, probably due to a retrotransposon insertion close to the alternative splicing site. In some other wild relatives of wheat, such as Aegilops comosa, there is only one copy of this gene, highly similar to WKS2, which is lucking the retrotransposon insertion. WKS2 gene present in wheat and WKS2-Ae from A. showed a different pattern of alternative splice variants, regardless of the presence of the retrotransposon insertion. Susceptible Bobwhite transformed with WKS2-Ae (without retrotansposon insertion in intron10), which derived from Aegilops comosaconferred resistance to stripe rust in wheat. The expression of WKS2-Ae in transgenic plants is up-regulated by temperature and pathogen infection. Combination of WKS1 and WKS2-Ae shows improved stripe rust resistance in WKS1×WKS2-Ae F1 hybrid plants. The obtained results show that WKS1 protein is accelerating programmed cell death observed in the regions surrounding a stripe rust infection in the wheat plants carrying the natural or transgenic WKS1 gene. Furthermore, characterization of the epistatic interactions of Yr36 and Yr18 demonstrated that these two genes have additive effects and can therefore be combined to increase partial resistance to this devastating pathogen of wheat. These achievements may have a broad impact on wheat breeding efforts attempting to protect wheat yields against one of the most devastating wheat pathogen.
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Avni, Adi, et Kirankumar S. Mysore. Functional Genomics Approach to Identify Signaling Components Involved in Defense Responses Induced by the Ethylene Inducing Xyalanase Elicitor. United States Department of Agriculture, décembre 2009. http://dx.doi.org/10.32747/2009.7697100.bard.

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Plant-microbe interactions involve a large number of global regulatory systems, which are essential for plants to protect themselves against pathogen attack. An ethylene-inducing xylanase (EIX) of Trichoderma viride is a potent elicitor of plant defense responses, like hypersensitive response (HR), in specific cultivars of tobacco (Nicotiana tabacum) and tomato (Lycopersicon esculentum). The central goal of this proposal was to investigate the molecular mechanisms that allow plants to specifically activate defense responses after EIX treatment. We proposed to identify cellular signaling components involved in the induction of HR by the EIX elicitor. The molecular genetic analysis of the signal transduction pathway that modulates hypersensitive responses is an important step in understanding the induction of plant defense responses. The genes that mediate LeEIX2-EIX dependent activation of resistance mechanisms remain to be identified. We used two approaches to identify the cellular signaling components that induce HR mediated by the EIX elicitor. In the first approach, we performed a yeast two-hybrid screening using LeEix2 as bait to identify plant proteins that interact with it. In the second approach, we used virus-induced gene silencing (VIGS) for a high-throughput screen to identify genes that are required for the induction of LeEIX2-EIX mediated HR. VIGS will also be used for functional characterization of genes that will be identified during the yeast two-hybrid screen. This investigation will shed light on cellular processes and signaling components involved in induction of general plant defense against pathogens and will provide the basis for future biotechnological approaches to improve plant resistance to pathogens. Several genes were indentified by the two approaches. We used the VIGS and yeast two hybrid approaches to confirm that activity of the genes initially identified by different procedure. Two genes inhibit the induction of HR by the fungal elicitor in the different systems; Tobacco-Harpin binding protein 1 and cyclopropyl isomerase.
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