Relevant bibliographies by topics / Promotor <Genetik>

Academic literature on the topic 'Promotor <Genetik>'

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Published: 26 July 2024

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Journal articles on the topic "Promotor <Genetik>"

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Oktariana, Desi, Irsan Saleh, Zen Hafy, and Iche Andriyani Liberty. "Peran Polimorfisme Promotor Gen Interleukin-10 pada Penyakit Kusta: Tinjauan Sistematik." Journal Of The Indonesian Medical Association 73, no. 1 (May 5, 2023): 15–24. http://dx.doi.org/10.47830/jinma-vol.73.1-2023-816.

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Pendahuluan: Kusta masih merupakan masalah penting di Indonesia karena jumlah kasus yang terus bertambah dan belum tereliminasi hingga saat ini. Salah satu sitokin yang berperan dalam kusta adalah interleukin-10 (IL-10), yang memiliki efek dominan anti-inflamasi dan mempengaruhi imunopatogenesis kusta. Faktor genetik, termasuk polimorfisme promotor gen IL-10, mempengaruhi jumlah IL-10 yang dihasilkan dan juga berkontribusi pada risiko terjadinya kusta. Oleh karena itu, penelitian ini bertujuan untuk memperoleh informasi tentang peran polimorfisme promotor gen IL-10 dalam kusta.Metode: Dalam penulisan ini, digunakan metode tinjauan sistematik yang bertujuan untuk mengidentifikasi, mengevaluasi, dan membahas secara mendalam karyakarya yang telah dipublikasikan melalui jurnal nasional dan internasional seperti Google Scholar, Science Direct, Elsevier, EBSCO, Medline, PubMed, Proquest, dan Wiley. Penelitian ini dilakukan dengan melakukan pencarian terhadap database tersebut menggunakan kata kunci yang relevan dengan topik penelitian. Setelah itu, dilakukan seleksi artikel berdasarkan kriteria inklusi dan eksklusi yang telah ditetapkan. Artikel yang memenuhi kriteria tersebut kemudian dievaluasi secara kualitas untuk memastikan keakuratan dan keandalan informasi yang disajikan. Akhirnya, hasil tinjauan sistematik ini digunakan untuk memperoleh informasi yang akurat dan terkini tentang peran polimorfisme promotor gen IL-10 dalam kusta.Hasil: Sebanyak 14 artikel dikaji dalam tinjauan sistematik ini dan didapatkan hasil yang beragam pada setiap populasi penelitian. Namun, terdapat kecenderungan bahwa polimorfisme promotor gen IL-10 terkait dengan penyakit kusta, yang ditemukan pada sebagian besar populasi penelitian.Kesimpulan: Polimorfisme promotor gen IL-10 cenderung terkait dengan penyakit kusta.
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Wurlina, Wurlina, Rimayanti Rimayanti, Mas'ud Hariadi, and Dewa Ketut Meles. "PEMBIBITAN DAN PENGGEMUKAN KAMBING “LOKETAWA” PENGHASIL DAGING DAN SUSU RAKITAN TEKNOBREEDING DAN TEKNOFATTENING PAKAN TANPA HIJAUAN (COMPLETE FEED)." Jurnal Layanan Masyarakat (Journal of Public Services) 1, no. 1 (June 1, 2017): 46. http://dx.doi.org/10.20473/jlm.v1i1.2017.46-50.

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IbM to business gorup in the goat breeding and fattening “Loketawa” aims to: 1) improve local goat genetics through IB using Etawa stud 2) obtain mother goats lust together done synchronization lusts 3) get mother goat bunting from once a year to 2 times a year 4) increase the number of children from 1 birth to 3-4 births using superovulation techniques 5) reduce production costs, make feed without forage and growth promotor. The method of implementation used 1) introducing stud Etawa meat and milk producers 2) synchronization lust using PGF2α 3) superovulation using hormone FSH and LH 4) IB on goats using Etawa goat cuttings, 5) the processing of feed without forage and growth promoter. The results are as follows: 1) as many as 20 heads of goats are simultaneously using 100% PGF2α, artificial insemination on 10 heads of goats without super ovulation has an average child of 1.6 births, while artificial insemination on 10 goats with super ovulation having an average child 3 births, 3) an average goat weight increase of 252.35 grams per day. AbstrakIpteks bagi Masyarakat yang dilakukan pada kelompok usaha pembibitan dan penggemukan kambing “Loketawa” bertujuan: 1) memperbaiki genetik kambing lokal melalui IB menggunakan pejantan Etawa 2) mendapatkan induk kambing birahi bersamaam dilakukan sinkronisasi birahi 3) mendapatkan induk kambing bunting dari setahun sekali menjadi 2 kali setahun 4) meningkatkan jumlah anak dari 1 ekor sekelahiran menjadi 3-4 ekor sekelahiran menggunakan teknik superovulasi 5) menekan biaya produksi, membuat pakan tanpa hijauan dan growth promotor. Metode pelaksanaan yang digunakan adalah 1) memperkenalkan pejantan Etawa penghasil daging dan susu 2) sinkronisasi birahi menggunakan PGF2α 3) superovulasi menggunakan hormon FSH dan LH 4) IB pada kambing menggunakan semen kambing Etawa, 5) pengolahan pakan tanpa hijauan dan growth promoter. Hasilnya adalah sebagai berikut : 1) sebanyak 20 ekor induk kambing mengalami birahi bersamaan menggunakan PGF2α sebesar 100%, Inseminasi buatan pada 10 ekor induk kambing tanpa super ovulasi mempunyai anak rata-rata 1,6 ekor sekelahiran, sedangkan Inseminasi buatan pada 10 ekor induk kambing dengan super ovulasi mempunyai anak rata-rata 3 ekor sekelahiran, 3) peningkatan berat badan kambing rata- rata 252,35 gram/ekor perhari.
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Li, Jinyang, Sheng Tong, Farrukh Raza Amin, Habiba Khalid, Kai Chen, Xiaoguang Zhao, Jinling Cai, and Demao Li. "Enhancing the Activity of a Self-Inducible Promoter in Escherichia coli through Saturation Mutation and High-Throughput Screening." Fermentation 9, no. 5 (May 13, 2023): 468. http://dx.doi.org/10.3390/fermentation9050468.

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The use of self-inducible promoters is a promising strategy to address metabolic imbalances caused by overexpression. However, the low activity of natural self-inducible promoters hinders their widespread application. To overcome this limitation, we selected the fic promoter as a model promoter to create an enhanced self-inducible promoter library using saturation mutations and high-throughput screening. Sequence analysis revealed that these promoters share certain characteristics, including semi-conservation in the −35 hexamer, highly conserved cytosine in the −17 motif (compared to −13 for other promoters), and moderate A+T content between positions −33 and −18 in the spacer region. Additionally, the discriminator region of these promotors features high A+T content in the first five bases. We identified PficI-17, PficII-33, and PficIII-14 promoters as the optional promoters in the −35 hexamer, spacer region, and discriminator mutation libraries, respectively. These promotors were used as representatives to measure the specific fluorescence and OD600 nm dynamics in different media and to confirm their effect on the expression of different proteins, including egfp (enhanced green fluorescence protein) and rfp (red fluorescence protein). Overall, our findings provide valuable guidance for modifying promoters and developing a promoter library suitable for regulating target genes.
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Shujaat, Muhammad, Abdul Wahab, Hilal Tayara, and Kil To Chong. "pcPromoter-CNN: A CNN-Based Prediction and Classification of Promoters." Genes 11, no. 12 (December 21, 2020): 1529. http://dx.doi.org/10.3390/genes11121529.

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A promoter is a small region within the DNA structure that has an important role in initiating transcription of a specific gene in the genome. Different types of promoters are recognized by their different functions. Due to the importance of promoter functions, computational tools for the prediction and classification of a promoter are highly desired. Promoters resemble each other; therefore, their precise classification is an important challenge. In this study, we propose a convolutional neural network (CNN)-based tool, the pcPromoter-CNN, for application in the prediction of promotors and their classification into subclasses σ70, σ54, σ38, σ32, σ28 and σ24. This CNN-based tool uses a one-hot encoding scheme for promoter classification. The tools architecture was trained and tested on a benchmark dataset. To evaluate its classification performance, we used four evaluation metrics. The model exhibited notable improvement over that of existing state-of-the-art tools.
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Mahoney, Michael E., and Daniel L. Wulff. "Mutations that Improve the pRE Promoter of Coliphage Lambda." Genetics 115, no. 4 (April 1, 1987): 591–95. http://dx.doi.org/10.1093/genetics/115.4.591.

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ABSTRACT The dya5 mutation, a C→T change at position -43 of the λ pRE promoter, results in a twofold increase in pRE activity in vivo. Smaller increases in pRE activity are found for the dya2 mutation, a T→C change at position -1 of pRE, and the dya3 mutation, an A→G change at +5 of pRE· The mutant pRE promoters retain complete dependence on cII protein for activity. These observations argue, at least for pRE-like promoters, that promoter activities are influenced by nucleotide sequences at least eight nucleotides to the 5′-side of the conventional -35 region consensus sequence, and by nucleotide sequences near the start-site of transcription. Although Hawley and McClure (1983) found A·T pairs more frequently than G·TC pairs in the region of -40 to -45 of prokaryotic promoters, other mutations that change a G·TC pair to an A·T pair at positions -41, -44 and -45 of pRE do not result in increased promoter activity. We also found that a T→C change at position -42 results in a mild decrease in promoter activity. These observations argue that Ts at positions -42 and -43 of pRE are required for maximum promoter activity, but do not support the hypothesis that As and Ts in the -40 to -45 region generally lead to higher promoter activities.
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George, Janet A., and Mary-Lou Pardue. "The Promoter of the Heterochromatic Drosophila Telomeric Retrotransposon, HeT-A, Is Active When Moved Into Euchromatic Locations." Genetics 163, no. 2 (February 1, 2003): 625–35. http://dx.doi.org/10.1093/genetics/163.2.625.

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Abstract The Drosophila telomeric retrotransposon, HeT-A, is found only in heterochromatin; therefore, its promoter must function in this chromatin environment. Studies of position effect variegation suggest that promoters of heterochromatic genes are very different from euchromatic promoters, but this idea has not been tested with isolated promoter sequences. The HeT-A promoter is the first heterochromatin promoter to be isolated and it is of interest to investigate its activity when removed from telomeric heterochromatin. This promoter was initially characterized by testing reporter constructs in transient transfection of cultured cells, an environment that may approximate its endogenous heterochromatin. We now report P-element-mediated transpositions of these constructs, testing the function of different parts of the putative promoter in euchromatin. Expression of endogenous HeT-A RNA shows marked developmental regulation and accumulates preferentially in replicating diploid tissues. HeT-A promoter constructs are active in all euchromatic locations tested and some display aspects of endogenous HeT-A stage- and cell-type expression programs. The activity of each promoter construct in euchromatic locations is also generally consistent with its activity in the transient transfection tests; a possibly significant exception is one sequence segment that appreciably enhanced activity in transient transfection but repressed promoter activity in euchromatin.
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Voelkel-Meiman, K., and G. S. Roeder. "Gene conversion tracts stimulated by HOT1-promoted transcription are long and continuous." Genetics 126, no. 4 (December 1, 1990): 851–67. http://dx.doi.org/10.1093/genetics/126.4.851.

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Abstract The recombination-stimulating sequence, HOT1, corresponds to the promoter of transcription by yeast RNA polymerase I. The effect of HOT1 on mitotic interchromosomal recombination was examined in diploid strains carrying a heterozygous URA3 gene on chromosome III. The frequency of Ura- recombinants was increased 20-fold when HOT1 was inserted into the chromosome III copy marked with URA3, at a location 48 kbp centromere-proximal to URA3. Ura- recombinants were increased only 2-fold when HOT1 and URA3 were on opposite homologues. These results suggest that most HOT1-promoted Ura- recombinants result from gene conversion and that sequences on the HOT1-containing chromosome are preferentially converted. Characterization of Ura- recombinants isolated from strains carrying multiple markers on chromosome III indicates that HOT1-promoted gene conversion tracts are unusually long (often greater than 75 kbp) and almost always continuous. Furthermore, conversion tracts frequently extend to both sides of HOT1. We suggest that HOT1 promotes the formation of a double-strand break which is often followed by exonucleolytic digestion. Repair of the broken chromosome could then result from gap repair or from replicative repair primed only by the centromere-containing chromosomal fragment.
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Davey, James A., and Corey J. Wilson. "Engineered signal-coupled inducible promoters: measuring the apparent RNA-polymerase resource budget." Nucleic Acids Research 48, no. 17 (September 5, 2020): 9995–10012. http://dx.doi.org/10.1093/nar/gkaa734.

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Abstract Inducible promoters are a central regulatory component in synthetic biology, metabolic engineering, and protein production for laboratory and commercial uses. Many of these applications utilize two or more exogenous promoters, imposing a currently unquantifiable metabolic burden on the living system. Here, we engineered a collection of inducible promoters (regulated by LacI-based transcription factors) that maximize the free-state of endogenous RNA polymerase (RNAP). We leveraged this collection of inducible promotors to construct simple two-channel logical controls that enabled us to measure metabolic burden – as it relates to RNAP resource partitioning. The two-channel genetic circuits utilized sets of signal-coupled transcription factors that regulate cognate inducible promoters in a coordinated logical fashion. With this fundamental genetic architecture, we evaluated the performance of each inducible promoter as discrete operations, and as coupled systems to evaluate and quantify the effects of resource partitioning. Obtaining the ability to systematically and accurately measure the apparent RNA-polymerase resource budget will enable researchers to design more robust genetic circuits, with significantly higher fidelity. Moreover, this study presents a workflow that can be used to better understand how living systems adapt RNAP resources, via the complementary pairing of constitutive and regulated promoters that vary in strength.
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Fandl, J. P., L. K. Thorner, and S. W. Artz. "Mutations that affect transcription and cyclic AMP-CRP regulation of the adenylate cyclase gene (cya) of Salmonella typhimurium." Genetics 125, no. 4 (August 1, 1990): 719–27. http://dx.doi.org/10.1093/genetics/125.4.719.

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Abstract We studied the expression of the cya promoter(s) in cya-lac fusion strains of Salmonella typhimurium and demonstrated cAMP receptor protein (CRP)-dependent repression by cAMP. Expression of cya was reduced about fourfold in cultures grown in acetate minimal medium as compared to cultures grown in glucose-6-phosphate minimal medium. Expression of cya was also reduced about fourfold by addition of 5 mM cAMP to cultures grown in glucose minimal medium. We constructed in vitro deletion and insertion mutations altering a major cya promoter (P2) and a putative CRP binding site overlapping P2. These mutations were recombined into the chromosome by allele replacement with M13mp::cya recombinant phages and the regulation of the mutant promoters was analyzed. A 4-bp deletion of the CRP binding site and a 4-bp insertion in this site nearly eliminated repression by cAMP. A mutant with the P2 promoter and the CRP binding site both deleted exhibited an 80% reduction in cya expression; the 20% residual expression was insensitive to cAMP repression. This mutant retained a Cya+ phenotype. Taken together, the results establish that the cya gene is transcribed from multiple promoters one of which, P2, is negatively regulated by the cAMP-CRP complex. Correction for the contribution to transcription by the cAMP-CRP nonregulated cya promoters indicates that the P2 promoter is repressed at least eightfold by cAMP-CRP.
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Greener, A., S. M. Lehman, and D. R. Helinski. "Promoters of the broad host range plasmid RK2: analysis of transcription (initiation) in five species of gram-negative bacteria." Genetics 130, no. 1 (January 1, 1992): 27–36. http://dx.doi.org/10.1093/genetics/130.1.27.

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Abstract A broad host range cloning vector was constructed, suitable for monitoring promoter activity in diverse Gram-negative bacteria. This vector, derived from plasmid RSF1010, utilized the firefly luciferase gene as the reporter, since the assay for its bioluminescent product is sensitive, and measurements can be made without background from the host. Twelve DNA fragments with promoter activity were obtained from broad host range plasmid RK2 and inserted into the RSF1010 derived vector. The relative luciferase activities were determined for these fragments in five species of Gram-negative bacteria. In addition, four promoters were analyzed by primer extension to locate transcriptional start sites in each host. The results show that several of the promoters vary substantially in relative strengths or utilize different transcriptional start sites in different bacteria. Other promoters exhibited similar activities and identical start sites in the five hosts examined.
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Dissertations / Theses on the topic "Promotor <Genetik>"

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Quehl, Eike. "Etablierung transgener Zelllinien zur Visualisierung der Aktivität des Doublecortin-Promotors als Modell der Neurogenese in vitro." kostenfrei, 2008. http://www.opus-bayern.de/uni-regensburg/volltexte/2009/1149/.

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Hohenstatt, Antonia. "Hypoxia inducible factor 1 (HIF 1) alpha- und beta-vermittelte Induktion der ABCA1-Promotor-Aktivität." kostenfrei, 2009. http://www.opus-bayern.de/uni-regensburg/volltexte/2009/1306/.

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Sommer, Heide. "Der Übergang von der Latenz zur lytischen Replikation des Epstein-Barr-Virus vergleichende Analysen zur Bedeutung regulatorischer HI-Motive im Promotor des viralen Gens BZLF-1 /." [S.l.] : [s.n.], 2001. http://deposit.ddb.de/cgi-bin/dokserv?idn=962765007.

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Sigvardsson, Mikael. "Regulation of immunoglobulin transcription during B-cell differentiation." Lund : Lund University, 1995. http://books.google.com/books?id=TJNqAAAAMAAJ.

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Ezpeleta, Jessica. "The characterization of the ABF-1 promoter." Scholarly Commons, 2001. https://scholarlycommons.pacific.edu/uop_etds/559.

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The basic helix-loop-helix (bHLH) family oftranscription factors consists of proteins involved in cellular proliferation and differentiation. The HLH structure plays a key role in protein-protein dimerization and with the DNA target sites, referred to as E boxes containing the consensus DNA sequence CANNTG. One class of mammalian class I bHLH proteins includes products of the E2A gene, which result from alternative splicing (E12, E47, and ITF), E2-2 and HEB. E2A proteins have also been detected in most cell lines with high levels of expression in lymphoid- and pancreatic cells. It has also been demonstrated that E2A is required for B cell maturation, T cell development and has been shown to function as tumor suppressors. To date, an E2A-interacting bHLH transcription factor largely restricted to activated B lymphocytes, called ABF -1, was isolated using the two-hybrid system. ABF -1 is the only B cell restricted bHLH protein isolated. ABF-1/E2A heterodimers have been detected in B lymphocytes. In these studies, the mapping of the ABF-1 promoter and the critical 5' regulatory elements that control ABF-1 gene expression were analyzed through 5' deletional analysis. 5' -DNA flanking pieces of the promoter region were created through PCR and inserted into a promoterless cloning vector containing the firefly luciferase reporter gene. RT -PCR analysis and anchored PCR was utilized to demonstrate the transcriptional activity of the - promoter region of the ABF-1 gene. Transient transfections were completed to determine critical regulatory sequences. The promoter location was confirmed through computer analysis of the nucleotide sequence and deletional analysis.
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Conradie, E. C. (Elizabeth Cornelia). "Promotor engineering in Saccharomyces cerevisiae for transcriptional control under different physiological conditions." Thesis, Stellenbosch : University of Stellenbosch, 2011. http://hdl.handle.net/10019.1/16512.

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Dissertation (PhD)--University of Stellenbosch, 2005.
ENGLISH ABSTRACT: To manipulate recombinant microorganisms for industrial processes, controllable genetic systems are needed that can coordinate expression of recombinant metabolic pathways. All components are sensitive to change and thus putative targets for modification and genetic elements and regulatory systems need to be understood and determined. Central in gene regulation is the transcription activators that mediate gene transcription mechanisms by binding to promoters in response to environmental signals. Promoter engineering entails the modification of transcription factors and their target promoters. In this study, a metabolic control system in Saccharomyces cerevisiae was constructed that would allow induction in response to physiological environment, specifically hypoxia and low temperature conditions. Two approaches were undertaken to find such a system. Firstly, a bi-directional reporter gene cloning vector was designed to search for novel hypoxiainducible promoters. Secondly, a transcription regulatory circuit was built, consisting of an inducible transcription regulator and promoter with a reporter gene through which it mediates transcription. Advantage was taken of the modular nature of proteins and functional domains originating from different transcriptional proteins were combined. A search for promoter elements sensitive to hypoxia from a S. cerevisiae genomic DNA (gDNA) library, using a bi-directional cloning vector, did not yield highly inducible promoters. It was concluded that a multitude of signals overlap, rendering genetic induction difficult to control. A synthetic regulatory system would minimize the impact of these multiple interactions. Such a genetic circuit was constructed, consisting of a chimeric transcription activator and a target fusion promoter. The chimeric transcription activator consisted of the GAL4 DNA binding domain, ADR1 TADIII transactivation domain and three domains of the MGA2 regulatory protein. The functional domains of Mga2p responsible for unregulated expression (at high basal levels) under both aerobic and hypoxia conditions were located, as well as a further upregulation under low temperature, and were mapped to the Nterminal and mid-Mga2p regions. A target fusion promoter consisting of a partial GAL10/1 promoter sequence and a Trichoderma reesei core xyn2 promoter were constructed as target for this chimeric transactivator. This synthetic promoter was fused to the T. reesei xyn2 open reading frame encoding for a readily assayable β-xylanase activity. Both the chimeric transactivator and fusion promoter-reporter gene cassettes were expressed from the same episomal plasmid, named pAR. Transformed into S. cerevisiae Y294, this regulatory system induced transcription under aerobic and hypoxia conditions. Furthermore, the reporter gene expression was upregulated by the chimeric transactivator at low temperatures. The chimeric transactivator mediated a seven-fold induction of the reporter gene under aerobic conditions in S. cerevisiae Y294 when transformed with plasmid AR. A two- to three-fold induction at 23ºC was reported under anaerobic conditions, relative to a reference strain expressing a transcription activator without the Mga2p domains. At 30ºC, a two- to three-fold induction under aerobic conditions and similar induction under oxygen-limited conditions were observed. Replacing the reporter gene with your favorite gene (for example a recombinant enzyme) and incorporating such a pAR system into a recombinant yeast should induce expression of the chosen gene under low temperatures, both aerobic and anaerobically (thus creating a controllable system). The system also has wider application in identifying other transcription factors’ signal-sensitive domains. The design of this system provides the ability to add a linker to a transactivator and to either create specific signal sensitivity or relieve the regulator of its signal dependence. It creates an easy system for assessing other transactivators and their domains with unknown functions and thus provides a ”workhorse and prospector in one”.
AFRIKAANSE OPSOMMING: Vir die manipulering van rekombinante mikroörganismes vir industriële prosesse word beheerbare genetiese stelsels benodig om gekoördineerde uitdrukking van rekombinante metaboliese weë teweeg te bring. Alle komponente van sulke stelsels is sensitief vir verandering en genetiese elemente en reguleerbare sisteme moet dus deeglik verstaan of bepaal word. Sentraal tot geenregulering is die transkripsie-aktiveerders wat geentranskripsie beheer deur aan promoters te bind in reaksie op eksterne omgewingsfaktore. Promotoringenieurswese behels wysigings van transkripsiefaktore en hul teikenpromotors. In hierdie studie is 'n genetiese beheerstelsel vir Saccaromyces cerevisiae ontwikkel wat induksie in reaksie tot spesifieke fisiologiese omgewingreaksies, naamlik hipoksie- en lae temperatuur, toelaat. Twee benaderings is gevolg: eerstens is ‘n tweerigting verklikker-geen vektor ontwikkel en gebruik om vir unieke induseerbare hipoksie-promoters te soek. Tweedens is ‘n transkripsie reguleringstelsel gebou wat uit ‘n induseerbare transkripsiereguleerder and promotor met ‘n verklikkergeen bestaan, waardeur transkripsie bemiddel kan word. Hierdie benadering benut die modulêre onderbou van proteïene en funksionele domeine afkomstig vanaf verskillende transkripsiefaktore is gekombineer. 'n Soektog na hipoksie-sensitiewe promotors vanuit 'n Saccharomyces cerevisiae-genoom- DNA (gDNA), deur van ‘n tweerigting verklikker-vektor gebruik te maak, het ongelukkig nie hoogs-induseerbare promotors opgelewer nie. Die gevolgtrekking was dat ‘n veelvoud van seine met mekaar oorvleuel en die beheer van genetiese induksie dus bemoeilik. Die ontwikkeling van ‘n sintetiese regulering-sisteem kan die impak van die veelvuldige interaksies verminder. Vir dié doel is ‘n sintetiese reguleringstelsel ontwerp, bestaande uit ‘n chimeriese transkripsie-aktiveerder met ‘n teiken fusie-promotor. Die chimeriese transaktiveerder bestaan uit die GAL4 DNA bindingsdomein, die ADR1 TAD III transaktiveringsdomein en drie domeine van die Mga2 reguleringsproteïen. In die studie is die funksionele domeins van Mga2p betrokke by lae temperatuur-respons en ongereguleerde uitdrukking (teen hoë basale vlakke) onder beide aërobiese en anaërobiese toestande aangedui en is tot die N-terminaal en middel-Mga2p areas gekarteer. ‘n Teiken-fusie-promoter, bestaande uit 'n gedeeltelike GAL1/10 DNA promotoropeenvolging en ‘n Trichoderma reesei kern xyn2-promoter, is as teiken vir hierdie chimeriese transaktiveerder saamgestel. Hierdie sintetiese promotor is aan die T. reesei xyn2 oopleesraam, wat vir ‘n maklik meetbare β-xylanase aktiwiteit kodeer, gekoppel. Beide die chimeriese transaktiveerder and fusie-promoter-verklikker-geenkaset word vanaf dieselfde episomale plasmied, bekend as pAR, uitgedruk. Hierdie reguleringsisteem induseer transkripsie onder aërobiese en hipoksie toestande in S. cerevisiae Y294. Verder word die verklikkergeen se uitdrukking deur die chimeriese transaktiveerder by lae temperature verhoog. Die chimeriese transaktiveerder induseer ‘n sewe-voudige induksie van die verklikkergeen onder aërobiese toestande by 23ºC vanaf die pAR-stelsel in S. cerevisiae Y294. ‘n Twee- tot drie-voudige induksie teen 23ºC is onder hipoksie toestande gevind, relatief tot induksievlakke van ‘n verwysingstam met ‘n transaktiveerder sonder die Mga2 domeine. By 30ºC is ‘n twee- tot drie-voudige induksie onder aërobiese en lae suurstofvlakke waargeneem. Deur die verklikker geen met ‘n jou-gunsteling-geen te vervang (bv. ‘n rekombinante ensiem) en so 'n pAR-sisteem in ‘n rekombinante gis te inkorporeer, word uitdrukking onder lae temperature onder beide aërobiese- en anaërobiese toestande geïnduseer (en sodoende word ‘n reguleerbare sisteem geskep). Die sisteem het wyer toepassing om sein-sensitiewe domeine van ander transkripsiefaktore te identifiseer. Die ontwerp van die stelsel maak dit moontlik om 'n skakel tot die transaktiveerder by te voeg wat óf sensitiwiteit tot 'n spesifieke sein skep, óf die reguleerder vanaf seinafhanklikheid verlos. So word ‘n bruikbare stelsel vir die bestudering van ander transaktivators en hul domeine met onbekende funksie geskep – ‘n “werksesel en prospekteerder in een”.
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Grade, Carla Vermeulen Carvalho 1983. "Caracterização funcional do promotor gênico da miostatina." [s.n.], 2013. http://repositorio.unicamp.br/jspui/handle/REPOSIP/317671.

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Orientador: Lucia Elvira Alvares
Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: A Miostatina é um regulador negativo da deposição de musculatura esquelética e mutações no gene que codifica esta proteína têm sido associadas a um aumento marcante na massa muscular de organismos vertebrados, resultado de hiperplasia e hipertrofia das fibras musculares. Nosso grupo identificou previamente o promotor basal do gene da Miostatina e análises de bioinformática revelaram a presença de sítios evolutivamente conservados para a ligação de CREB, Meis, FXR e NFY, além de um sítio TATA. No presente trabalho nós utilizamos mutagênese sítio-dirigida para gerar diversas construções delecionais que possuem um ou mais sítios mutados, e testamos sua atividade in vitro usando mioblastos C2C12 de camundongo sob condições de proliferação e diferenciação, para analisar o papel destes sítios de ligação sobre a atividade do promotor. Os resultados mostraram que FXR aparentemente não confere efeito na atividade transcricional do promotor da Miostatina em ambos os momentos analisados, indicando que o papel regulador desta proteína pode estar relacionado ao controle da expressão da Miostatina em outro tipo celular, que não o mioblasto. O NFY apresentou um papel de ativador transcricional, enquanto CREB e Meis atuaram inicialmente como repressores durante a proliferação, passando a relaxar esta repressão durante a diferenciação dos mioblastos, permitindo que a atividade do promotor aumentasse significativamente. Trabalhando juntos, estes fatores de transcrição são capazes de manter a atividade do promotor em níveis mais baixos durante a proliferação dos mioblastos e, com o início da diferenciação, a repressão é liberada, e os níveis de atividade podem aumentar. Este padrão está de acordo com o padrão de expressão dinâmico observado para a proteína da Miostatina durante o desenvolvimento da musculatura esquelética em vertebrados
Abstract: Myostatin is a negative regulator of skeletal muscle deposition and mutations in the gene that encodes this protein have been associated to a remarkable increase in skeletal muscle mass, attributable to both hyperplasia and hypertrophy. We have previously identified Myostatin's basal promoter and bioinformatic analyses revealed the presence of evolutionarily conserved binding sites for CREB, Meis, FXR and NFY, besides a TATA box. In the present study we used site-directed mutagenesis to generate several expression constructs possessing one or more mutated sites, and tested their activity in vitro using mouse C2C12 myoblasts in proliferation and differentiation conditions, to analyze the role of these sites on the activity of the promoter. The results show that FXR appears not to confer any effect on the transcriptional activity of the promoter in both conditions, indicating that the regulatory role of this protein might be involved in the control of Myostatin expression in another cell type. NFY presents a role as transcriptional activator, while CREB and Meis act initially as repressors during proliferation, releasing this repression upon differentiation, which allows the activity of the promoter to significantly increase. Working together, these transcription factors are capable of maintaining the promoter activity at lower levels during the proliferation of myoblasts and, upon differentiation, the repression is released, and activity levels can be increased. This pattern is in agreement with the dynamic expression pattern observed for Myostatin during the skeletal muscle development in vertebrates
Doutorado
Histologia
Doutor em Biologia Celular e Estrutural
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Zhang, Xiao-Qun. "Functional Studies on the PDGFR α gene promoter and effects of autocrine PDGF-A stimulation in vivo." Doctoral thesis, Uppsala universitet, Institutionen för genetik och patologi, 2001. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-1455.

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Platelet-derived growth factor receptor α (PDGFRα) plays an important role during embryogenesis. After implantation, the patterns of expression of Pdgfrα and its ligand Pdgf-A undergo an "autocrine-paracrine transition", in that Pdgf-A becomes expressed in the ectoderm and epithelia, while Pdgfrα is expressed in the adjacent mesenchymal tissue. In human tumors, such as malignant glioma, both PDGF and PDGFRα are overexpressed within the same tissue, indicating that an autocrine PDGF loop is generated in the tumors. This thesis is focused on the in vivo functionality of the PDGFRα gene (PDGFRA) promoter, arid on the effect of autocrine PDGF-A stimulation in transgenic n-iice during embryogenesis. To test the in vivo promoter function of a human PDGFRA 2.2 kb 5' flanking fragment, we generated transgenic mouse lines and found that the 2.2 kb fragment was able to promote lacZ reporter gene expression in most of the endogenous Pdgfra expressing tissues. Absence of expression and "ectopic" expression of the transgenic lacZ were also observed. To investigate the autocrine PDGF effect, we produced autocrine PDGF-As (A short-chain) transient transgenic embryos. These transgenic embryos carried a 6 kb mouse Pdgfra 5' flanking sequence linked to a human PDGF-As cDNA. The pattern of expression of the PDGF-As transgene mRNA was similar to that of lacZ. Some of the transgenic embryos exhibited severe abnormal phenotypes, such as midline fusion defects in the cephalic and craniofacial region and small body size, and these embryos die at mid-gestation stage. These findings indicate that a paracrine pattern of expression and the dosage of PDGF are important for sustaining normal embryo development, especially with regard to the middline fusion in craniofacial regions. The possible signaling pathways that may be involved in regulating Pdgfra activity were also studied by comparison of patterns of mRNA expression of Gli, Ptc, and Paxl with that of Pdgfra. The results pointed to the possibility that the Shh signaling pathway may be involved in the regulation of Pdgfra expression for example during early bone and foregut development. The specific regulatory mechanisms may vary for different tissues.
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Warshamana, Gnana Sakuntala. "Interactions of T7 RNA polymerase with its promoters : Part I: T7 promoter contacts essential for promoter activity in vivo ; Part II: Isolation and characterization of a mutant T7 RNA polymerase with altered promoter specificity." Diss., Georgia Institute of Technology, 1992. http://hdl.handle.net/1853/26303.

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Lorson, Christian. "An analysis of transcriptional regulation of the MVM capsid gene promoter." free to MU campus, to others for purchase, 1997. http://wwwlib.umi.com/cr/mo/fullcit?p9841319.

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Books on the topic "Promotor <Genetik>"

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Lutz, Nover, ed. Plant promoters and transcription factors. Berlin: Springer-Verlag, 1994.

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Lawrence, Privalsky Martin, ed. Transcriptional corepressors: Mediators of eukaryotic gene repression. Berlin: Springer, 2001.

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International Congress and Exhibition on Nutrition, Fitness, and Health (2006 Shanghai, China). Nutrition and fitness: Cultural, genetic, and metabolic aspects. Edited by Simopoulos Artemis P. 1933-. Basel: Karger, 2008.

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Ponnambalam, Sreenivasan. Transcription initiation at the 'Escherichia Coli' galactose Operon promoter region: Genetic and biochemicalstudies. Birmingham: University of Birmingham, 1988.

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Yeomans, Gary M. Generic milk promotion in the United States and Canada. Uckfield: Nuffield Farming Scholarships Trust, 1998.

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EPA Workshop on the Development of Risk Assessment Methodologies for Tumor Promoters (1987 Bethesda, Md.). Report of the EPA Workshop on the development of risk assessment methodologies for tumor promoters. Washington, DC: Office of Health and Environmental Assessment and Office of Regulatory Support and Scientific Analysis, Office of Research and Development, U.S. Environmental Protection Agency, 1987.

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Kamp, Philip Vande. Commodity promotion programs in the United States. Ithaca, N.Y: Dept. of Agricultural, Resource, and Managerial Economics, College of Agriculture and Life Sciences, Cornell University, 1999.

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Kaiser, Harry Mason. An analysis of generic dairy promotion in the United States. Ithaca, N.Y: Dept. of Agricultural, Resource, and Managerial Economics, College of Agriculture and Life Sciences, Cornell University, 1995.

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Blisard, William Noel. Generic promotion of agricultural products: Balancing producers' and consumers' needs. [Washington, D.C.?]: U.S. Dept. of Agriculture, Economic Research Service, 1989.

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Blisard, William Noel. Generic promotion of agricultural products: Balancing producers' and consumers' needs. [Washington, D.C.?]: U.S. Dept. of Agriculture, Economic Research Service, 1989.

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Book chapters on the topic "Promotor <Genetik>"

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Komarnytsky, Slavko, and Nikolai Borisjuk. "Functional Analysis of Promoter Elements in Plants." In Genetic Engineering, 113–41. Boston, MA: Springer US, 2003. http://dx.doi.org/10.1007/978-1-4615-0073-5_6.

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Guilfoyle, Tom J. "The Structure of Plant Gene Promoters." In Genetic Engineering, 15–47. Boston, MA: Springer US, 1997. http://dx.doi.org/10.1007/978-1-4615-5925-2_2.

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Pelham, Hugh. "Properties and Uses of Heat Shock Promoters." In Genetic Engineering, 27–44. Boston, MA: Springer US, 1987. http://dx.doi.org/10.1007/978-1-4684-5377-5_2.

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Santos, Efrén, Ricardo Pacheco, Liliana Villao, Luis Galarza, Daniel Ochoa, Carlos Jordán, and José Flores. "Promoter Analysis in Banana." In Banana: Genomics and Transgenic Approaches for Genetic Improvement, 157–79. Singapore: Springer Singapore, 2016. http://dx.doi.org/10.1007/978-981-10-1585-4_11.

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Reuter, Ingmar, Thomas Werner, and Edgar Wingender. "Computer-Assisted Methods for the Identification and Characterization of Polymerase II Promoters." In Genetic Engineering, 25–40. Boston, MA: Springer US, 1998. http://dx.doi.org/10.1007/978-1-4899-1739-3_2.

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Nowell, Peter C. "Genetic Instability and Tumor Development." In Boundaries between Promotion and Progression during Carcinogenesis, 221–31. Boston, MA: Springer US, 1991. http://dx.doi.org/10.1007/978-1-4684-5994-4_19.

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Angel, J. M., and J. DiGiovanni. "Genetics of Skin Tumor Promotion." In Animal Models of Cancer Predisposition Syndromes, 143–57. Basel: KARGER, 1999. http://dx.doi.org/10.1159/000062010.

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Barry, Margaret M. "A Generic Template for Implementing Mental Health Promotion." In Implementing Mental Health Promotion, 131–59. Cham: Springer International Publishing, 2019. http://dx.doi.org/10.1007/978-3-030-23455-3_5.

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Howard, Daniel, and Karl Benson. "Evolutionary Computation Method for Promoter Site Prediction in DNA." In Genetic and Evolutionary Computation — GECCO 2003, 1690–701. Berlin, Heidelberg: Springer Berlin Heidelberg, 2003. http://dx.doi.org/10.1007/3-540-45110-2_62.

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Nordgren, Anders. "The Human Genome Project: Justification, Promotion, and Access to Results." In Responsible Genetics, 91–126. Dordrecht: Springer Netherlands, 2001. http://dx.doi.org/10.1007/978-94-015-9741-8_3.

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Conference papers on the topic "Promotor <Genetik>"

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Frénoy, Antonine, François Taddei, and Dusan Misevic. "Constrained Genetic Architecture Promotes Cooperation." In Artificial Life 14: International Conference on the Synthesis and Simulation of Living Systems. The MIT Press, 2014. http://dx.doi.org/10.7551/978-0-262-32621-6-ch004.

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Frénoy, Antonine, François Taddei, and Dusan Misevic. "Constrained Genetic Architecture Promotes Cooperation." In Artificial Life 14: International Conference on the Synthesis and Simulation of Living Systems. The MIT Press, 2014. http://dx.doi.org/10.1162/978-0-262-32621-6-ch004.

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Medvet, Eric, Alberto Bartoli, and Giovanni Squillero. "An effective diversity promotion mechanism in grammatical evolution." In GECCO '17: Genetic and Evolutionary Computation Conference. New York, NY, USA: ACM, 2017. http://dx.doi.org/10.1145/3067695.3076057.

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Talamini, Jacopo, Giovanni Scaini, Eric Medvet, and Alberto Bartoli. "Selfish vs. global behavior promotion in car controller evolution." In GECCO '18: Genetic and Evolutionary Computation Conference. New York, NY, USA: ACM, 2018. http://dx.doi.org/10.1145/3205651.3208254.

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Singh, Abhyudai, Cesar A. Vargas, and Rajesh Karmakar. "Stochastic analysis of genetic promoter architectures with memory." In 2013 IEEE 52nd Annual Conference on Decision and Control (CDC). IEEE, 2013. http://dx.doi.org/10.1109/cdc.2013.6761034.

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Byeon, B., and K. Rasheed. "Bayesian Networks and Genetic Algorithms for Promoter Recognition." In IASTED Technology Conferences 2010. Calgary,AB,Canada: ACTAPRESS, 2010. http://dx.doi.org/10.2316/p.2010.728-030.

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Larson, Ari, Anton Bernatskiy, Collin Cappelle, Ken Livingston, Nicholas Livingston, John Long, Jodi Schwarz, Marc Smith, and Josh Bongard. "Recombination Hotspots Promote the Evolvability of Modular Systems." In GECCO '16: Genetic and Evolutionary Computation Conference. New York, NY, USA: ACM, 2016. http://dx.doi.org/10.1145/2908961.2908990.

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Carvalho, Jonata Tyska, and Stefano Nolfi. "Exploiting environmental differentiation to promote evolvability in artificial evolution." In GECCO '17: Genetic and Evolutionary Computation Conference. New York, NY, USA: ACM, 2017. http://dx.doi.org/10.1145/3067695.3075994.

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Guzman-Ruiz, Omar, Manuel Mejia-Lavalle, Alicia Martinez, and Yasmin Hernandez. "Machine learning Algorithms applied to Genetic Promoter Sequences Recognition." In 2020 International Conference on Mechatronics, Electronics and Automotive Engineering (ICMEAE). IEEE, 2020. http://dx.doi.org/10.1109/icmeae51770.2020.00016.

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"Analysis of the activity of the DR5 promoter in tuber-forming plants." In Plant Genetics, Genomics, Bioinformatics, and Biotechnology. Novosibirsk ICG SB RAS 2021, 2021. http://dx.doi.org/10.18699/plantgen2021-111.

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Reports on the topic "Promotor <Genetik>"

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Brinckerhoff, Constance E. Genetic Analysis of a Single Nucleotide Polymorphism in the Matrix Metalloproteinase 1 Promoter in Breast Cancer. Fort Belvoir, VA: Defense Technical Information Center, July 2002. http://dx.doi.org/10.21236/ada407580.

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Brinckerhoff, Constqance B. Genetic Analysis of a Single Nucleotide Polymorphism in the Matrix Metalloproteinase 1 Promoter in Breast Cancer. Fort Belvoir, VA: Defense Technical Information Center, July 2003. http://dx.doi.org/10.21236/ada419338.

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Strauss, S. H., V. Busov, K. Kosola, J. Kennedy, J. Morrell, C. Ma, A. Elias, and E. Etherington. Genetic modification of gibberellic acid signaling to promote carbon sequestration in tree roots and stems. Office of Scientific and Technical Information (OSTI), May 2009. http://dx.doi.org/10.2172/952484.

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Busov, Victor. GENETIC MODIFICATION OF GIBBERELLIC ACID SIGNALING TO PROMOTE CARBON SEQUESTRATION IN TREE ROOTS AND STEMS. Office of Scientific and Technical Information (OSTI), March 2013. http://dx.doi.org/10.2172/1067341.

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Norelli, John L., Moshe Flaishman, Herb Aldwinckle, and David Gidoni. Regulated expression of site-specific DNA recombination for precision genetic engineering of apple. United States Department of Agriculture, March 2005. http://dx.doi.org/10.32747/2005.7587214.bard.

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Objectives: The original objectives of this project were to: 1) evaluate inducible promoters for the expression of recombinase in apple (USDA-ARS); 2) develop alternative selectable markers for use in apple to facilitate the positive selection of gene excision by recombinase (Cornell University); 3) compare the activity of three different recombinase systems (Cre/lox, FLP/FRT, and R/RS)in apple using a rapid transient assay (ARO); and 4) evaluate the use of recombinase systems in apple using the best promoters, selectable markers and recombinase systems identified in 1, 2 and 3 above (Collaboratively). Objective 2 was revised from the development alternative selectable markers, to the development of a marker-free selection system for apple. This change in approach was taken due to the inefficiency of the alternative markers initially evaluated in apple, phosphomannose-isomerase and 2-deoxyglucose-6-phosphate phosphatase, and the regulatory advantages of a marker-free system. Objective 3 was revised to focus primarily on the FLP/FRT recombinase system, due to the initial success obtained with this recombinase system. Based upon cooperation between researchers (see Achievements below), research to evaluate the use of the FLP recombinase system under light-inducible expression in apple was then conducted at the ARO (Objective 4). Background: Genomic research and genetic engineering have tremendous potential to enhance crop performance, improve food quality and increase farm profits. However, implementing the knowledge of genomics through genetically engineered fruit crops has many hurdles to be overcome before it can become a reality in the orchard. Among the most important hurdles are consumer concerns regarding the safety of transgenics and the impact this may have on marketing. The goal of this project was to develop plant transformation technologies to mitigate these concerns. Major achievements: Our results indicate activity of the FLP\FRTsite-specific recombination system for the first time in apple, and additionally, we show light- inducible activation of the recombinase in trees. Initial selection of apple transformation events is conducted under dark conditions, and tissue cultures are then moved to light conditions to promote marker excision and plant development. As trees are perennial and - cross-fertilization is not practical, the light-induced FLP-mediated recombination approach shown here provides an alternative to previously reported chemically induced recombinase approaches. In addition, a method was developed to transform apple without the use of herbicide or antibiotic resistance marker genes (marker free). Both light and chemically inducible promoters were developed to allow controlled gene expression in fruit crops. Implications: The research supported by this grant has demonstrated the feasibility of "marker excision" and "marker free" transformation technologies in apple. The use of these safer technologies for the genetic enhancement of apple varieties and rootstocks for various traits will serve to mitigate many of the consumer and environmental concerns facing the commercialization of these improved varieties.
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Jung, Carina, Karl Indest, Matthew Carr, Richard Lance, Lyndsay Carrigee, and Kayla Clark. Properties and detectability of rogue synthetic biology (SynBio) products in complex matrices. Engineer Research and Development Center (U.S.), September 2022. http://dx.doi.org/10.21079/11681/45345.

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Synthetic biology (SynBio) aims to rationally engineer or modify traits of an organism or integrate the behaviors of multiple organisms into a singular functional organism through advanced genetic engineering techniques. One objective of this research was to determine the environmental persistence of engineered DNA in the environment. To accomplish this goal, the environmental persistence of legacy engineered DNA building blocks were targeted that laid the foundation for SynBio product development and application giving rise to “post-use products.” These building blocks include genetic constructs such as cloning and expression vectors, promoter/terminator elements, selectable markers, reporter genes, and multi-cloning sites. Shotgun sequencing of total DNA from water samples of pristine sites was performed and resultant sequence data mined for frequency of legacy recombinant DNA signatures. Another objective was to understand the fate of a standardized contemporary synthetic genetic construct (SC) in the context of various chassis systems/genetic configurations representing different degrees of “genetic bioavailability” to the environmental landscape. These studies were carried out using microcosms representing different environmental matrices (soils, waters, wastewater treatment plant (WWTP) liquor) and employed a novel genetic reporter system based on volatile organic compounds (VOC) detection to assess proliferation and persistence of the SC in the matrix over time.
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Solvin, Thomas, and Inger Sundheim Fløistad. Statistics: Forest Seeds and Plants in the Nordic Region – Version 2023. The Nordic Genetic Resource Center (NordGen), August 2023. http://dx.doi.org/10.53780/qoub7866.

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The Nordic Genetic Resource Center (NordGen) is the joint genebank and knowledge center for genetic resources in the Nordic countries. Our mission is to conserve and promote the sustainable use of genetic diversity among animals, forests and plants that are important for Nordic agriculture and forestry. “Statistics: Forest Seeds and Plants in the Nordic Region – Version 2023” is the second edition in a biennial statistics report on forest seed and plant material in the Nordic countries. The first edition was published in 2021. This edition has been expanded by including more statistics and more species than the first report, as well as including more recent data from the years 2020 and 2021. The report compiles statistics and reports contributed by representatives of each country in the NordGen Forest Regeneration Council.
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Dawson, William O., and Moshe Bar-Joseph. Creating an Ally from an Adversary: Genetic Manipulation of Citrus Tristeza. United States Department of Agriculture, January 2004. http://dx.doi.org/10.32747/2004.7586540.bard.

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Citrus is one of the major agricultural crops common to Israel and the United States, important in terms of nutrition, foreign exchange, and employment. The economy of both citrus industries have been chronically plagued by diseases caused by Citrus tristeza virus (CTV). The short term solution until virus-resistant plants can be used is the use of mild strain cross-protection. We are custom designing "ideal" protecting viruses to immunize trees against severe isolates of CTV by purposely inoculating existing endangered trees and new plantings to be propagated as infected (protected) citrus budwood. We crossed the substantial technological hurdles necessary to accomplish this task which included developing an infectious cDNA clone which allows in vitro manipulation of the virus and methods to then infect citrus plants. We created a series of hybrids between decline-inducing and mild CTV strains, tested them in protoplasts, and are amplifying them to inoculate citrus trees for evaluation and mapping of disease determinants. We also extended this developed technology to begin engineering transient expression vectors based on CTV as tools for genetic improvement of tree crops, in this case citrus. Because of the long periods between genetic transformation and the ultimate assay of mature tree characteristics, there is a great need for an effective system that allows the expression or suppression of target genes in fruiting plants. Virus-based vectors will greatly expedite progress in citrus genetic improvement. We characterized several components of the virus that provides necessary information for designing virus-based vectors. We characterized the requirements of the 3 ’-nontranslated replication promoter and two 3 ’-ORF subgenomic (sg) mRNA controller elements. We discovered a novel type of 5’-terminal sgRNAs and characterized the cis-acting control element that also functions as a strong promoter of a 3 ’-sgRNA. We showed that the p23 gene controls negative-stranded RNA synthesis and expression of 3 ’ genes. We identified which genes are required for infection of plants, which are host range determinants, and which are not needed for plant infection. We continued the characterization of native dRNA populations and showed the presence of five different classes including class III dRNAs that consists of infectious and self-replicating molecules and class V dRNAs that contain all of the 3 ’ ORFs, along with class IV dRNAs that retain non-contiguous internal sequences. We have constructed and tested in protoplasts a series of expression vectors that will be described in this proposal.
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Fridman, Eyal, and Eran Pichersky. Tomato Natural Insecticides: Elucidation of the Complex Pathway of Methylketone Biosynthesis. United States Department of Agriculture, December 2009. http://dx.doi.org/10.32747/2009.7696543.bard.

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Plant species synthesize a multitude of specialized compounds 10 help ward off pests. and these in turn may well serve as an alternative to synthetic pesticides to reduce environmental damage and health risks to humans. The general goal of this research was to perform a genetic and biochemical dissection of the natural-insecticides methylketone pathway that is specific to the glandular trichomes of the wild species of tomato, Solanumhabrochaites f. glabratum (accession PI126449). Previous study conducted by us have demonstrated that these compounds are synthesized de novo as a derivate pathway of the fatty acid biosynthesis, and that a key enzyme. designated MethylketoneSynthase 1 (MKS 1). catalyzes conversion of the intermediate B-ketoacyl- ACPs to the corresponding Cn-1 methylketones. The approach taken in this proposed project was to use an interspecific F2 population. derived from the cross between the cultivated lV182 and the wild species PIl26449. for three objectives: (i) Analyze the association between allelic status of candidate genes from the fatty acid biosynthesis pathway with the methylketone content in the leaves (ii) Perform bulk segregant analysis of genetic markers along the tomato genome for identifying genomic regions that harbor QTLs for 2TD content (iii) Apply differential gene expression analysis using the isolated glands of bulk segregant for identifying new genes that are involved in the pathway. The genetic mapping in the interspecific F2 population included app. 60 genetic markers, including the candidate genes from the FAS pathway and SSR markers spread evenly across the genome. This initial; screening identified 5 loci associated with MK content including the candidate genes MKS1, ACC and MaCoA:ACP trans. Interesting observation in this genetic analysis was the connection between shape and content of the glands, i.e. the globularity of the four cells, typical to the wild species. was associated with increased MK in the segregating population. In the next step of the research transcriptomic analysis of trichomes from high- and 10w-MK plants was conducted. This analysis identified a new gene, Methy1ketone synthase 2 (MKS2), whose protein product share sequence similarity to the thioesterase super family of hot-dog enzymes. Genetic analysis in the segregating population confirmed its association with MK content, as well as its overexpression in E. coli that led to formation of MK in the media. There are several conclusions drawn from this research project: (i) the genetic control of MK accumulation in the trichomes is composed of biochemical components in the FAS pathway and its vicinity (MKS 1 and MKS2). as well as genetic factors that mediate the morphology of these specialized cells. (ii) the biochemical pathway is now realized different from what was hypothesized before with MKS2 working upstream to I\1KS 1 and serves as the interface between primary (fatty acids) and secondary (MK) metabolism. We are currently testing the possible physical interactions between these two proteins in vitro after the genetic analysis showed clear epistatic interactions. (iii) the regulation of the pathway that lead to specialized metabolism in the wild species is largely mediated by transcription and one of the achievements of this project is that we were able to isolate and verify the specificity of the MKS1 promoter to the trichomes which allows manipulation of the pathways in these cells (currently in progress). The scientific implications of this research project is the advancement in our knowledge of hitherto unknown biochemical pathway in plants and new leads for studying a new family in plants (hot dog thioesterase). The agricultural and biotechnological implication are : (i) generation of new genetic markers that could assist in importing this pathway to cultivated tomato hence enhancing its natural resistance to insecticides, (ii) the discovery of MKS2 adds a new gene for genetic engineering of plants for making new fatty acid derived compounds. This could be assisted with the use of the isolated and verified MKS1 promoter. The results of this research were summarized to a manuscript that was published in Plant Physiology (cover paper). to a chapter in a proceeding book. and one patent was submitted in the US.
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Zhang, Hongbin, Shahal Abbo, Weidong Chen, Amir Sherman, Dani Shtienberg, and Frederick Muehlbauer. Integrative Physical and Genetic Mapping of the Chickpea Genome for Fine Mapping and Analysis of Agronomic Traits. United States Department of Agriculture, March 2010. http://dx.doi.org/10.32747/2010.7592122.bard.

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Chickpea is the third most important pulse crop in the world and ranks first in the Middle East; however, it has been subjected to only limited research in modern genomics. In the first period of this project (US-3034-98R) we constructed two large-insert BAC and BIBAC libraries, developed 325 SSR markers and mapped QTLs controlling ascochyta blight resistance (ABR) and days to first flower (DTF). Nevertheless, the utilities of these tools and results in gene discovery and marker-assisted breeding are limited due to the absence of an essential platform. The goals of this period of the project were to use the resources and tools developed in the first period of the project to develop a BAC/BIBAC physical map for chickpea and using it to identify BAC/BIBACcontigs containing agronomic genes of interest, with an emphasis on ABR and DTF, and develop DNA markers suitable for marker-assisted breeding. Toward these goals, we proposed: 1) Fingerprint ~50,000 (10x) BACs from the BAC and BIBAC libraries, assemble the clones into a genome-wide BAC/BIBAC physical map, and integrate the BAC/BIBAC map with the existing chickpea genetic maps (Zhang, USA); 2) fine-map ABR and DTFQTLs and enhance molecular tools for chickpea genetics and breeding (Shahal, Sherman and DaniShtienberg, Israel; Chen and Muehlbauer; USA); and 3) integrate the BAC/BIBAC map with the existing chickpea genetic maps (Sherman, Israel; Zhang and Chen, USA). For these objectives, a total of $460,000 was requested originally, but a total of $300,000 was awarded to the project. We first developed two new BAC and BIBAC libraries, Chickpea-CME and Chickpea- CHV. The chickpea-CMEBAC library contains 22,272 clones, with an average insert size of 130 kb and equivalent to 4.0 fold of the chickpea genome. The chickpea-CHVBIBAC library contains 38,400 clones, with an average insert size of 140 kb and equivalent to 7.5 fold of the chickpea genome. The two new libraries (11.5 x), along with the two BAC (Chickpea-CHI) and BIBAC (Chickpea-CBV) libraries (7.1 x) constructed in the first period of the project, provide libraries essential for chickpea genome physical mapping and many other genomics researches. Using these four libraries we then developed the proposed BAC/BIBAC physical map of chickpea. A total of 67,584 clones were fingerprinted, and 64,211 (~11.6 x) of the fingerprints validated and used in the physical map assembly. The physical map consists of 1,945 BAC/BIBACcontigs, with each containing an average of 39.2 clones and having an average physical length of 559 kb. The contigs collectively span ~1,088 Mb, being 1.49 fold of the 740- Mb chickpea genome. Third, we integrated the physical map with the two existing chickpea genetic maps using a total of 172 (124 + 48) SSR markers. Fourth, we identified tightly linked markers for ABR-QTL1, increased marker density at ABR-QTL2 and studied the genetic basis of resistance to pod abortion, a major problem in the east Mediterranean, caused by heat stress. Finally, we, using the integrated map, isolated the BAC/BIBACcontigs containing or closely linked to QTL4.1, QTL4.2 and QTL8 for ABR and QTL8 for DTF. The integrated BAC/BIBAC map resulted from the project will provide a powerful platform and tools essential for many aspects of advanced genomics and genetics research of this crop and related species. These includes, but are not limited to, targeted development of SNP, InDel and SSR markers, high-resolution mapping of the chickpea genome and its agronomic genes and QTLs, sequencing and decoding of all genes of the genome using the next-generation sequencing technology, and comparative genome analysis of chickpea versus other legumes. The DNA markers and BAC/BIBACcontigs containing or closely linked to ABR and DTF provide essential tools to develop SSR and SNP markers well-suited for marker-assisted breeding of the traits and clone their corresponding genes. The development of the tools and knowledge will thus promote enhanced and substantial genetic improvement of the crop and related legumes.
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