Dissertationen zum Thema „Genetics engineering“
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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.
Der volle Inhalt der QuelleENGLISH 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”.
Wilsher, Julie Ann. „Protein engineering of chymosin“. Thesis, Birkbeck (University of London), 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.300804.
Der volle Inhalt der QuelleHill, Philip John. „PCR based gene engineering“. Thesis, University of Nottingham, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.317040.
Der volle Inhalt der QuelleSutherland, John David. „Genetic engineering of penicillin biosynthesis“. Thesis, University of Oxford, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.253132.
Der volle Inhalt der QuelleHilyard, Katherine L. „Protein engineering of antibody combining sites“. Thesis, University of Oxford, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.291278.
Der volle Inhalt der Quelle黃雅誼 und Nga-yi Queenie Wong. „DNA engineering utilizing thymidylate synthase A (thyA) selection system in Escherichia coli“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2001. http://hub.hku.hk/bib/B31226851.
Der volle Inhalt der QuelleZainuddin. „Genetic transformation of wheat (Triticum aestivum L.)“. Title page, Contents and Abstract only, 2000. http://web4.library.adelaide.edu.au/theses/09APSP/09apspz21.pdf.
Der volle Inhalt der Quelleau, dweston@ncwa com, und Delys Eleanor Weston. „Democracy and political economy of genetic engineering“. Murdoch University, 2007. http://wwwlib.murdoch.edu.au/adt/browse/view/adt-MU20070327.143205.
Der volle Inhalt der QuelleMacKenzie, Donald J. „Molecular characterization of potato virus S and genetic engineering of virus resistant plants“. Thesis, University of British Columbia, 1990. http://hdl.handle.net/2429/30622.
Der volle Inhalt der QuelleMedicine, Faculty of
Biochemistry and Molecular Biology, Department of
Graduate
Purohit, Shri Kant. „Analysis of nodulin-44 gene of soybean“. Thesis, McGill University, 1987. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=66088.
Der volle Inhalt der QuelleMauro, Vincent Peter. „Structure and regulation of nodulin genes of soybean“. Thesis, McGill University, 1986. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=75360.
Der volle Inhalt der QuelleForghani, Farnaz. „Protein engineering of bacteriophage Mu transposase“. Thesis, McGill University, 1990. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=60444.
Der volle Inhalt der QuelleRichards, James Edward. „Engineering a helper virus-free reverse genetics system for rotavirus“. Thesis, University of Cambridge, 2012. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.610743.
Der volle Inhalt der QuelleTorres, Arzayus Maria Isabel. „Engineering yam mosaic virus resistance in Nicotiana benthamiana using genetic transformation techniques“. Thesis, Imperial College London, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.264199.
Der volle Inhalt der QuelleChirakkal, Haridasan. „Enzyme engineering of chloramphenicol acetyltransferase by genetic selection and site-directed mutagenesis“. Thesis, University of Sheffield, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.267199.
Der volle Inhalt der QuelleTaylor, Noah David. „Engineering of Allosteric Transcription Factors and Their Use for Metabolic Pathway Evolution“. Thesis, Harvard University, 2016. http://nrs.harvard.edu/urn-3:HUL.InstRepos:26718755.
Der volle Inhalt der QuelleMedical Sciences
Samaan, L. Z. „Comparative studies using Agrobacterium spp. as vectors for genetic engineering of higher plants“. Thesis, University of Nottingham, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.373321.
Der volle Inhalt der QuellePrakash, Satya. „De novo engineering of trans-activating riboswitches in E. coli“. Thesis, University of Warwick, 2018. http://wrap.warwick.ac.uk/114021/.
Der volle Inhalt der QuelleWilliamson, Phillip C. „A Novel Mechanism for Site-Directed Mutagenesis of Large Catabolic Plasmids Using Natural Transformation“. Thesis, University of North Texas, 2001. https://digital.library.unt.edu/ark:/67531/metadc2828/.
Der volle Inhalt der QuelleRios, Villanueva Xavier. „Toward Multiplex Genome Engineering in Mammalian Cells“. Thesis, Harvard University, 2013. http://dissertations.umi.com/gsas.harvard:11179.
Der volle Inhalt der QuelleUnderhill, Michele F. „Engineering mRNA translation initiation in Chinese hamster ovary cells for enhanced production of recombinant proteins“. Thesis, University of Kent, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.322824.
Der volle Inhalt der QuelleSeah, Stephen Y. K. „Phenylalanine dehydrogenase of Bacillus sphaericus : kinetic characterisation and homology-based engineering of new substrate specificities“. Thesis, University of Sheffield, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.245580.
Der volle Inhalt der QuelleWu, Ying. „Protein engineering of human alpha 1-antitrypsin gene and the study of secretion defective mutants“. Thesis, University of Southampton, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.385366.
Der volle Inhalt der QuelleGanesan, Savita Ayre Brian Gordon. „FLP-mediated conditional loss of an essential gene to facilitate complementation assays“. [Denton, Tex.] : University of North Texas, 2007. http://digital.library.unt.edu/permalink/meta-dc-5180.
Der volle Inhalt der QuelleShareck, Julie. „Isolation and characterization of a cryptic plasmid from Lactobacillus plantarum“. Thesis, McGill University, 2005. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=84072.
Der volle Inhalt der QuelleAbioye, Jumai Adeola. „Engineering chimaeric recombinases for HIV-1 proviral DNA excision“. Thesis, University of Glasgow, 2018. http://theses.gla.ac.uk/9143/.
Der volle Inhalt der QuelleAzizi, Bahareh. „Chemical Complementation: A Genetic Selection System in Yeast for Drug Discovery, Protein Engineering, and for Deciphering and Assembling Biosynthetic Pathways“. Diss., Available online, Georgia Institute of Technology, 2005, 2005. http://etd.gatech.edu/theses/available/etd-07182005-102856/.
Der volle Inhalt der QuelleAllen M. Orville, Committee Member ; Sheldon W. May, Committee Member ; Jung H. Choi, Committee Member ; Mostafa A. El-Sayed, Committee Member ; Donald F. Doyle, Committee Chair.
Wells, Carol Dawn. „The functional significance of the G to A point mutation in the promoter region of the Apolipoprotein AI gene“. Master's thesis, University of Cape Town, 1993. http://hdl.handle.net/11427/27143.
Der volle Inhalt der QuelleRogers, Jameson Kerr. „Biosensing for Multiplexed Genome Engineering: Applications in Renewable Chemical Production“. Thesis, Harvard University, 2015. http://nrs.harvard.edu/urn-3:HUL.InstRepos:17467367.
Der volle Inhalt der QuelleEngineering and Applied Sciences - Engineering Sciences
Romano, Eduardo O. „Selection indices for combining marker genetic data and animal model information /“. This resource online, 1993. http://scholar.lib.vt.edu/theses/available/etd-09192009-040546/.
Der volle Inhalt der QuelleEdwards, Matthew Douglas. „Information-sharing models for computational genetics“. Thesis, Massachusetts Institute of Technology, 2016. http://hdl.handle.net/1721.1/105572.
Der volle Inhalt der QuelleThis electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections.
Cataloged from student-submitted PDF version of thesis.
Includes bibliographical references (pages 97-105).
Modern genetics has been transformed by a dramatic explosion of data. As sample sizes and the number of measured data types grow, the need for computational methods tailored to deal with these noisy and complex datasets increases. In this thesis, we develop and apply integrated computational and biological approaches for two genetic problems. First, we build a statistical model for genetic mapping using pooled sequencing, a powerful and efficient technique for rapidly unraveling the genetic basis of complex traits. Our approach explicitly models the pooling process and genetic parameters underlying the noisy observed data, and we use it to calculate accurate intervals that contain the targeted regions of interest. We show that our model outperforms simpler alternatives that do not use all available marker data in a principled way. We apply this model to study several phenotypes in yeast, including the genetic basis of the surprising phenomenon of strain-specific essential genes. We demonstrate the complex genetic basis of many of these strain-specific viability phenotypes and uncover the influence of an inherited virus in modifying their effects. Second, we design a statistical model that uses additional functional information describing large sets of genetic variants in order to predict which variants are likely to cause phenotypic changes. Our technique is able to learn complicated relationships between candidate features and can accommodate the additional noise introduced by training on groups of candidate variants, instead of single labeled variants. We apply this model to a large genetic mapping study in yeast by collecting multiple genome-wide functional measurements. By using our model, we demonstrate the importance of several molecular phenotypes in predicting genetic impact. The common themes in this thesis are the development of computational models that accurately reflect the underlying biological processes and the integration of carefully controlled biological experiments to test and utilize our new models.
by Matthew Douglas Edwards.
Ph. D.
Bekker, Tamrin Annelie. „Molekulere karakterisering van 'n Aegilops speltoides verhaalde translokasie en verkorte vorms“. Thesis, Stellenbosch : University of Stellenbosch, 2009. http://hdl.handle.net/10019.1/1854.
Der volle Inhalt der QuelleGene transfer from wild gras species to wheat is complicated by the simultaneous integration of large amounts of alien chromatin. The alien chromatin containing the target gene is inherited as a linkage block and the phenomenon is known as linkage drag. The degree of linkage drag depends on whether, and how readily, recombination occurs between the foreign and wheat chromatin. The S13 translocation line was developed by the department of Genetics, US. A cross was made between Chinese Spring and a leaf rust resistant Aegilops speltoides accession. Resistant backcross F1 was backcrossed to Chinese Spring and W84-17. S13 was selected from the backcross progeny and found to carry three rust resistance genes temporarily named LrS13, SrS13 and YrS13. Unfortunately, the resistance genes were completely linked to gametocidal (Gc) genes that were co-transferred from the wild parent. In wheat Gc genes cause reduced fertility, poor plant phenotype and hybrid necrosis. In order to use employ the rust resistance genes commercially they need to be separated from the Gc genes. At the onset of this study four putative shortened forms of the S13 translocation were provided. The four lines were identified in a homoeologous paring induction experiment (involving the test cross 04M127). This study aimed to achieve the following: (i) characterize the four recombinants with the use of molecular markers, (ii) use the knowledge gained to identify further recombinants in the 04M127 cross, (iii) identify the shortest (most useful) recombinant, and (iv) attempt to shorten the shortest recombinant form still further and thereby remove as many of the Gc genes as possible. In total, seven recombinants of the S13 translocation (04M127-1, -2, -3, -4, -7, -11 and -12; referred to as recombinant group A) were identified and characterised with microsatellite and SCAR markers. These recombinants have exchanged different amounts of foreign chromatin for wheat chromatin, but were still associated with Gc genes, showing hybrid necrosis and seed shrivelling. Some of the recombinants have lost the undesirable „brittle rachis‟ phenotype which occurs in Ae. speltoides and the S13 translocation line. In plants VII having this trait, the rachis spontaneously disarticulates after the third spikelet upon ripening of the ear. Recombinant 3 appeared to be least affected by Gc genes and was therefore used in further attempts to shorten the translocation. Recombinant 3 was crossed with wheat (W84-17) and resistant F1 (heterozygous for the translocation) were test crossed with Chinese Spring nullisomic 3A tetrasomic 3B/D plants. Thirty five resistant testcross F1 plants were identified (named recombinant group B). The resistant group B recombinants as well as nine susceptible test cross F1 (which also appeared to be recombinant) were characterised making use of microsatellites and a SCAR marker. From the results it appeared that each of the 35 resistant plants exchanged substantial amounts of Ae. speltoides chromatin for wheat chromatin. The species chromatin that remained (and which contains LrS13) is probably located either close to the 3AS telomere or within the proximal regions of 3AS and 3AL. A SCAR marker that has been developed specifically for the S13 translocation provided useful confirmation of the presence of Ae. speltoides chromatin in the 35 recombinants. If the SCAR marker proves to be tightly linked to LrS13 it may eventually be used for marker assisted selection of the resistance or it may be employed in continued attempts to reduce the amount of foreign chromatin. Seedling rust resistance tests showed that the recombinants have lost SrS13 and YrS1 during recombination. An attempt was also made to develop additional markers that specifically detect the translocation in order to further characterise the group B recombinants. Published information on Ae. speltoides specific repeated and transposon sequences were obtained and used for primer design. Unfortunately, no suitable markers could be found and the primers that were designed tended to amplify the same fragments in both the wheat and species genomes. DArT markers were also employed in an attempt to characterise the 35 group B recombinants and controls. The DArT results provided an independent verification of the results obtained with the microsatellite markers. The DArT results confirmed that the group B recombinants exchanged large amounts of species chromatin for wheat chromatin. Even though the 35 resistant group B recombinants have undergone extensive recombination they still show signs of residual Gc effects. It is believed these effects can be removed by continued backcrossing to wheat accompanied by selection against Gc symptoms. While the effects of Gc genes per se were not studied, their properties were reminiscent of those of transposable elements. Indications were that complex interactions involving the Gc genes themselves as well as genetic factors in the wheat genome may have a drastic effect on the selective survival of recombinant gametes.
Wang, Ming-Bo. „Isolation of phloem specific gene promoters for use in genetic engineering of insect resistance in rice“. Thesis, Durham University, 1994. http://etheses.dur.ac.uk/5146/.
Der volle Inhalt der QuelleKentner, Jeffrey Louis. „Engineering the zinc finger recombinase for use in targeted genomic editing“. Thesis, University of Glasgow, 2015. http://theses.gla.ac.uk/6910/.
Der volle Inhalt der QuelleBevington, Linda K. „The creation of humankind in the image of God and the incarnation of Christ implications for human genetic engineering, reproductive technology, and cloning /“. Theological Research Exchange Network (TREN), 1997. http://www.tren.com.
Der volle Inhalt der QuelleUrbach, Ena. „Evolution and population genetics of Prochlorococcus marinus“. Thesis, Massachusetts Institute of Technology, 1995. http://hdl.handle.net/1721.1/37755.
Der volle Inhalt der QuelleIncludes bibliographical references.
by Ena Urbach.
Ph.D.
Pauls, David G. „Evangelical attitudes towards human enhancement a survey of the Midwest District of the Evangelical Free Church of America /“. Theological Research Exchange Network (TREN) Theological Research Exchange Network (TREN) Access this title online, 2006. http://www.tren.com.
Der volle Inhalt der QuelleZhang, Ling 1962. „Molecular cloning and characterization of the chicken ornithine decarboxylase gene“. Thesis, McGill University, 1994. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=22831.
Der volle Inhalt der QuellePhuwadol, Bangrak Lily Eurwilaichitr. „Expression and secretion of giant catfish growth hormone in methylotrophic yeast pichia pastoris /“. Abstract, 1999. http://mulinet3.li.mahidol.ac.th/thesis/2542/42E-PhuwadolB.pdf.
Der volle Inhalt der QuelleMacfarlane, Hayley Louise. „Engineering site-specific recombinases for use in synthetic biology“. Thesis, University of Glasgow, 2017. http://theses.gla.ac.uk/8378/.
Der volle Inhalt der QuelleYang, Joyce Lichi. „Developments in Human Pluripotent Stem Cell Genome Engineering and in Situ Sequencing Technologies“. Thesis, Harvard University, 2015. http://nrs.harvard.edu/urn-3:HUL.InstRepos:17467524.
Der volle Inhalt der QuelleMedical Sciences
Butterfield, Michael Keith. „Marker assisted breeding in sugarcane : a complex polyploid“. Thesis, Link to the online version, 2007. http://hdl.handle.net/10019.1/1203.
Der volle Inhalt der QuelleChuang, William 1970. „Design of a genetics database for medical research“. Thesis, Massachusetts Institute of Technology, 2000. http://hdl.handle.net/1721.1/86291.
Der volle Inhalt der QuelleIncludes bibliographical references (leaves 54-57, first group).
by William Chuang.
S.B.and M.Eng.
Espach, Yolandi. „The detection of mycoviral sequences in grapevine using next-generation sequencing“. Thesis, Stellenbosch : Stellenbosch University, 2013. http://hdl.handle.net/10019.1/80025.
Der volle Inhalt der QuelleENGLISH ABSTRACT: Metagenomic studies that make use of next-generation sequencing (NGS) generate large amounts of sequence data, representing the genomes of multiple organisms of which no prior knowledge is necessarily available. In this study, a metagenomic NGS approach was used to detect multiple novel mycoviral sequences in grapevine phloem tissue. Individual sequencing libraries of doublestranded RNA (dsRNA) from two grapevine leafroll diseased (GLD) and three shiraz diseased (SD) vines were sequenced using an Illumina HiScanSQ instrument. Over 3.2 million reads were generated from each of the samples and these reads were trimmed and filtered for quality before being de novo assembled into longer contigs. The assembled contigs were subjected to BLAST (Basic Local Alignment Search Tool) analyses against the NCBI (National Centre for Biotechnology Information) database and classified according to database sequences with which they had the highest identity. Twenty-six putative mycovirus species were identified, belonging to the families Chrysoviridae, Endornaviridae, Narnaviridae, Partitiviridae and Totiviridae. Two of the identified mycoviruses, namely grapevine-associated chrysovirus (GaCV) and grapevine-associated mycovirus 1 (GaMV-1) have previously been identified in grapevine while the rest appeared to be novel mycoviruses not present in the NCBI database. Primers were designed from the de novo assembled mycoviral sequences and used to screen the grapevine dsRNA used for sequencing as well as endophytic fungi isolated from the five sample vines. Only two mycoviruses, related to sclerotinia sclerotiorum partitivirus S and chalara elegans endornavirus 1 (CeEV-1), could be detected in grapevine dsRNA and in fungus isolates. In order to validate the presence of mycoviruses in grapevine phloem tissue, two additional sequencing runs, using an Illumina HiScanSQ and an Applied Biosystems (ABI) SOLiD 5500xl instrument respectively, were performed. These runs generated more and higher quality sequence data than the first sequencing run. Twenty-two of the putative mycoviral sequences initially detected were detected in the subsequent sequence datasets, as well as an additional 29 species not identified in the first HiScanSQ sequence datasets. The samples harboured diverse mycovirus populations, with as many as 19 putative species identified in a single vine. This indicates that the complete virome of diseased grapevines will include a high number of mycoviruses. Additionally, the complete genome of a novel endornavirus, for which we propose the name grapevine endophyte endornavirus (GEEV), was assembled from one of the second HiScanSQ sequence datasets. This is the first complete genome of a mycovirus detected in grapevine. Grapevine endophyte endornavirus has the highest sequence similarity to CeEV-1 and is the same virus that was previously detected in fungus isolates using the mycovirus primers. The virus was detected in two fungus isolates, namely Stemphylium sp. and Aureobasidium pullulans, which is of interest since mycoviruses are not known to be naturally associated with two distinctly different fungus genera. Mycoviral sequence data generated in this study can be used to further investigate the diversity and the effect of mycoviruses in grapevine.
AFRIKAANSE OPSOMMING: Metagenomiese studies, wat gebruik maak van volgende-generasie volgordebepalingstegnologie, het die vermoë om die genetiese samestelling van veelvoudige onbekende organismes te bepaal deurdat dit groot hoeveelhede data genereer. Die bogenoemde tegniek was in hierdie studie aangewend om aantal nuwe mikovirusse in die floëem weefsel van wingerd te identifiseer. Dubbelstring-RNS was gesuiwer vanuit twee druiwestokke met rolbladsiekte en drie met shirazsiekte en Illumina HiScanSQ instrument is gebruik om meer as 3.2 miljoen volgorde fragmente te genereer van elk van die monsters. Lae-kwaliteit volgordes was verwyder en die oorblywende kort volgorde fragmente was saamgestel om langer konstrukte te vorm wat met behulp van BLAST soektogte teen die NCBI databasis geïdentifiseer kon word. Ses-en-twintig mikovirus spesies, wat aan die families Chrysoviridae, Endornaviridae, Narnaviridae, Partitiviridae en Totiviridae behoort, was geïdentifiseer. Twee van die geïdentifiseerde mikovirusse, naamlik grapevine-associated chrysovirus (GaCV) en grapevine-associated mycovirus 1 (GaMV-1), was voorheen al in wingerd gekry terwyl die res nuwe mikovirusse is wat tans nie in die NCBI databasis voorkom nie. Inleiers was ontwerp vanaf die saamgestelde mikovirus basisvolgordes en gebruik om wingerd dubbelstring-RNS sowel as swamme wat vanuit die wingerd geïsoleer is te toets vir die teenwoordigheid van hierdie mikovirusse. Slegs twee mikovirusse, wat onderskeidelik verwant is aan sclerotinia sclerotiorum partitivirus S en chalara elegans endornavirus 1 (CeEV-1), kon deur middel van die inleiers in wingerd en swam isolate geïdentifiseer word. Twee addisionele volgordebepalingsreaksies, wat gebruik gemaak het van die Illumina HiScanSQ en ABI SOLiD 5500xl volgordebepalingsplatforms, was gebruik om die teenwoordigheid van mikovirusse in wingerd te bevestig. Groter hoeveelheid volgorde fragmente was geprodusser wat ook van hoër gehalte was as dié van die eerste volgordebepalingsreaksie. Twee-en-twintig mikovirus spesies kon weer geïdentifiseer word, sowel as 29 spesies wat nie in die eerste HiScanSQ basisvolgorde datastelle gevind was nie. Die wingerdstokke wat in hierdie studie ondersoek was, het hoë diversiteit van mikovirusse bevat aangesien daar tot 19 mikovirus spesies in enkele wingerdstok geïdentifiseer was. Dit is aanduiding dat volledige virus profiele van siek wingerdstokke aantal mikovirusse sal insluit. Die vollengte genoomvolgorde van voorheen onbekende endornavirus was saamgestel vanuit een van die tweede HiScanSQ volgorde datastelle. Dit is die eerste mikovirus wat in wingerd gevind word waarvan die volledige genoomvolgorde bepaal is en ons stel die naam grapevine endophyte endornavirus (GEEV) voor vir hierdie virus. Grapevine endophyte endornavirus is die naaste verwant aan CeEV-1 en is dieselfde virus wat voorheen in wingerd dubbelstring-RNS en swam isolate gevind was deur middel van die mikovirus inleiers. Swam isolate waarin GEEV gevind is, was geïdentifiseer as Stemphylium sp. en Aureobasidium pullulans. Dit is van belang dat GEEV in twee swam isolate gevind is wat aan verskillende genusse behoort aangesien hierdie verskynsel nog nie voorheen in die natuur gevind is nie. Mikovirus nukleiensuurvolgordes wat in hierdie studie bepaal was kan gebruik word in toekomstige studies om die verskeidenheid en impak van mikovirusse in wingerd verder te ondersoek.
National Research Foundation (NRF)
Stellenbosch University
Nettelblad, Carl. „Using Markov models and a stochastic Lipschitz condition for genetic analyses“. Licentiate thesis, Uppsala universitet, Avdelningen för teknisk databehandling, 2010. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-120295.
Der volle Inhalt der QuelleeSSENCE
O'Kennedy, Martha Margaretha. „Genetic enhancement of pearl millet“. Thesis, Stellenbosch : Stellenbosch University, 2004. http://hdl.handle.net/10019.1/49974.
Der volle Inhalt der QuelleENGLISH ABSTRACT: The aim of this study was toe stablish a reliable protocol for the production 0 f transgenic pearl millet as this will open new avenues for augmenting the gene pool of this crop. This was achieved by identifying a highly regenerabie genotype and optimisation of a tissue culture system, and biolistic protocol f or stable integration of selected transgenes. Both a negative, herbicide resistance selectable marker gene, bar, and a positive selectable marker gene, manA, were individually introduced in order to identify and establish a reliable transformation protocol. The optimised transformation protocol was then used to introduce an antifungal gene in the genome of pearl millet to enhance resistance to the biotrophic fungus Sclerospora graminicola. S. graminicola, an obligate oomycetous fungal phytopathogen, is the causal agent of downy mildew in pearl millet plants and a major constraint in the production of pearl millet. A single component of antifungal resistance was introduced into the genome of pearl millet, as preliminary work towards determining its role in the total plant defence system. The approach chosen was to introduce a hydrolytic enzyme, 13-1,3- glucanase, from Trichoderma atroviride (formerly T. harzianum), a soil-borne filamentous fungus, capable of parasitizing several plant pathogenic fungi. It was anticipated that introducing this glucanase gene from T. atroviride which degrades glucan in the fungal cell walls, would significantly contribute to the improvement of resistance against downy mildew. Constructs were prepared containing the gene (gluc78) encoding a 78 kDa beta-1,3- glucanase. The constructs were prepared containing the gluc78 gene driven either by a strong constitutive promoter (ubiquitin promoter, exon and intron) or a wound inducible promoter, the potato proteinase inhibitor ilK gene promoter. The wound inducible promoter includes either an AMV leader' sequence or the rice Act1 intron to obtain higher expression levels in the monocotyledonous plant. The transformation efficiency using the particle inflow gun and the herbicide resistance gene, bar, was improved from 0.02% on a MS based medium, to 0.19 or 0.72% with manA as selectable marker gene on MS or L3 based medium, respectively. However, individual experiments, introducing manA as selectable marker gene, resulted in frequencies of 1.2 and 3%. This translated to one transformation event per plate, which contains on average 31-35 pre-cultured immature zygotic embryos. This is the first report of t he successful introduction and expression of a 13-1,3-glucanase encoding gene from a biocontrol fungus not only under constitutive expression but also under wound inducible expression in a plant. Optimisation of genetic engineering of pearl millet, a cereal crop recalcitrant to transformation, and the introduction of an antifungal transgene, was accomplished in this study. Initial results hint that expression of this transgene enhances resistance to S. graminicola.
AFRIKAANSE OPSOMMING: Die doel van die studie was om 'n betroubare genetiese transformeringsprotokol vir pêrel manna te ontwikkel. Hiervoor moes eerstens 'n regenereerbare genotipe geidentifiseer word. Twedens moes 'n betroubare weefselkultuur en biolistiese transformeringssisteem ontwikkel word. Beide die onkruiddoder bestandheidsgeen, bar, en 'n positiewe selektiewe geen, manA, is vir die doel van die projek onafhanklik in die genoom van pêrel manna in gekloneer. Die optimale sisteem is vervolgens aangewend om 'n geen wat potensieël verbeterde bestandheid teen die biotrofiese swam Sclerospora graminicola wat donsige meeldou by plante veroorsaak, in pêrel manna in te kloneer. 'n Enkele komponent van bestandheid is in die genetiese material van pêrel manna in gekloneer as inleidende werk om die rol van hierdie geen in die totale verdedigingsisteem te bepaal. Die benadering wat gekies was, behels die klonering van 'n hidrolitiese ensiem 13-1,3-glukanase, van Trichoderma atroviride (voorheen T. harzianum), 'n grondgedraagde swam, wat op 'n aantal ander plantpatogene fungus kan parasiteer. Die verwagting is dat klonering van hierdie 13- 1,3-glukanase geen van T. atroviride wat die glukaan verteer in die selwande van swamme, 'n groot verbetering tot die bestandheid teen donsige meeldou sal meebring. Konstrukte is voorberei wat die gluc78 geen bevat wat kodeer vir die 78 kDa beta-1,3-glukanase protein. Die konstrukte wat voorberei is bevat die gluc78 geen geinduseer deur of 'n sterk konstituwe promoter (ubiquitin promoter, exon en intron) of deur 'n wond geinduseerde promoter, die aartappel proteinase inhibeerder ilK geen promoter. Hierdie promoter word gevolg deur of 'n AMV leier volgorde of die rys Act1 intron om verhoogde uitdruk vlakke in monokotiele plante te verseker. As die partikel invloei geweer in kombinasie met die onkruiddoderbestandheidsgeen gebruik word, was die doeltreffendheid van transformasie 0.02% op 'n MS gebasseerde groeimedium. 'n Transformasie doeltreffendheid van onderskeidelik 0.19 en 0.72% is verkry wanneer die manA as selektiewe geen gebruik is op MS of L3 gebasseerde medium. Twee individual eksperimente, waar die manA geen as selektiewe geen gebruik is, het gelei tot 'n transformasie doeltreffendheid van 1.2 of 3%. Dit gee 'n gemiddelde van een transformasie per plaat wat 31 tot 35 voorafgekweekte onvolwasse embrios bevat. Hierdie is d ie eerste verslag van d ie suksesvolle klonering en uitdrukking van 'n 13-1,3-glukanasekoderende geen van 'n swam wat as 'n biologiese beheeragent gebruik word. Hierdie is nie alleenlik onder konstitutiewe uitdrukking nie, maar ook 0 nder wond g einduseerde u itdruk in' n p lant. In hierdie studie is die 0 ptimisering van genetiese verbetering van pêrel manna, 'n graan gewas wat gehard is teen transformasie, deur die klonering van 'n bestandheidsgeen in die genoom van hierdie gewas gedoen. Aanvanklike resultate dui daarop dat die uitdruk van hierdie geen lei tot verbeterde bestandheid teen S. graminicola.
Selgelid, Michael J. „Neugenics : genetically-informed reproductive decision making /“. Diss., Connect to a 24 p. preview or request complete full text in PDF format. Access restricted to UC IP addresses, 2001. http://wwwlib.umi.com/cr/ucsd/fullcit?p3015841.
Der volle Inhalt der QuelleLui, Jervis C. (Jervis Chun-Wai) 1973. „Development of a pedigree analysis tool for genetics counselors“. Thesis, Massachusetts Institute of Technology, 1998. http://hdl.handle.net/1721.1/47601.
Der volle Inhalt der QuelleIncludes bibliographical references (p. 50).
by Jervis C. Lui.
S.B.and M.Eng.
Boyle, Patrick M. „Network-Scale Engineering: Systems Approaches to Synthetic Biology“. Thesis, Harvard University, 2012. http://dissertations.umi.com/gsas.harvard:10298.
Der volle Inhalt der QuelleNdimande, Sandile. „Increasing cellulosic biomass in sugarcane“. Thesis, Stellenbosch : Stellenbosch University, 2014. http://hdl.handle.net/10019.1/86296.
Der volle Inhalt der QuelleENGLISH ABSTRACT: Increased demand of petroleum, declining fossil fuel reserves, geopolitical instability and the environmentally detrimental effects of fossil fuels have stimulated research to search for alternative sources of energy such as plant derived biofuels. The main feedstocks for production of first generation biofuels (bioethanol) are currently sucrose and starch, produced by crops such as sugarcane, sugarbeet, maize, and cassava. The use of food crop carbohydrates to produce biofuels is viewed as competing for limited agronomic resources and jeopardizing food security. Plants are also capable of storing sugars in their cell walls in the form of polysaccharides such as cellulose, hemicelluloses and pectin, however those are usually cross-linked with lignin, making their fermentation problematic, and are consequently referred to as lignocellulosics. Current technologies are not sufficient to degrade these cell wall sugars without large energy inputs, therefore making lignocellulosic biomass commercially unviable as a source of sugars for biofuel production. In the present study genes encoding for enzymes for cellulosic, hemicellulosic and starch-like polysaccharides biosynthesis were heterologously expressed to increase the amount of fermentable sugars in sugarcane. Transgenic lines heterologously expressing CsCesA, encoding a cellulose synthase from the marine invertebrate Ciona savignyi showed significant increases in their total cellulose synthase enzyme activity as well as the total cellulose content in internodal tissues. Elevation in cellulose contents was accompanied by a rise in hemicellulosic glucose content and uronic acid amounts, while total lignin was reduced in internodal tissues. Enzymatic saccharification of untreated lignocellulosic biomass of transgenic sugarcane lines had improved glucose release when exposed to cellulose hydrolyzing enzymes. Calli derived from transgenic sugarcane lines ectopically expressing galactomannan biosynthetic sequences ManS and GMGT from the cluster bean (Cyamopsis tetragonoloba) were observed to be capable of producing a galactomannan polysaccharide. However, after regeneration, transgenic sugarcane plants derived from those calli were unable to produce the polymer although the inserted genes were transcribed at the mRNA level. While the ectopic expression of Deinococcus radiodurans amylosucrase protein in the cytosol had a detrimental effect on the growth of transgenic lines (plants showed stunted growth through the 18 months growth period in greenhouse), contrastingly targeting the amylosucrase protein into the vacuole resulted in 3 months old transgenic lines which were having high maltooligosaccharide and soluble sugar (sucrose, glucose and fructose) levels in leaves. After 18 months growing in the greenhouse, the mature transgenic lines were morphologically similar to the untransformed lines and also contained comparable maltooligosaccharide and soluble sugar and starch amounts. The non-biosynthesis of galactomannan and amylose polysaccharides in the matured transgenic plants may be due to post-transcriptional protein processing and or protein instability, possibly explainable by other epigenetic mechanisms taking place to regulate gene expression in the at least allo-octaploid species of sugarcane under investigation in this study.