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Dissertations / Theses on the topic 'Chromosomes, Artificial, Bacterial'

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Published: 4 June 2021

Last updated: 7 February 2022

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1

Tonga, Lavon Paongo Hutt-Fletcher Lindsey M. "Development of an Akata-based bacterial artificial chromosome." Diss., UMK access, 2006.

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Thesis (Ph. D.)--School of Biological Sciences. University of Missouri--Kansas City, 2006.<br>"A dissertation in molecular biology and biochemistry and cell biology and biophysics." Advisor: Lindsey M. Hutt-Fletcher. Typescript. Vita. Title from "catalog record" of the print edition Description based on contents viewed Nov. 9, 2007. Includes bibliographical references (leaves 89-103). Online version of the print edition.

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2

Kong, Sim Yee. "Plasmid DNA and bacterial artificial chromosomes processing for gene therapy and vaccination : studies on membrane sterile filtration." Thesis, University College London (University of London), 2007. http://discovery.ucl.ac.uk/1444899/.

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Plasmids currently applied in clinical trials are generally < 20 kb, but the interest in larger circular vectors is rising. However, due to their size and character, filterability and irreversible damage intensified by elongational shear are major concerns during a sterilising filtration procedure. Therefore, key parameters affecting the normal-flow membrane filtration performance of solutions containing purified plasmid DNA and bacterial artificial chromosomes were investigated in this study. Two small scale filtration systems were designed to enable information on material properties to be obtained. Firstly, a pressure driven syringe system was used to conduct constant flux experiments. Data on transmission and degradation could be obtained rapidly and only required small sample volumes (< 1 mL). Secondly, a positive pressure filtration system which permitted operations at constant transmembrane pressure had been applied to determine the filter capacity, this information is useful to facilitate the scale-up of a membrane filtration process. The results showed transmission of DNA vectors decreased linearly with molecular size (6-1 16 kb) and confocal microscopy images confirmed that a fraction of the DNA molecules were being retained by the membrane filters. Degradation increased with molecular weight, flux and number of filtration passes for vectors > 20 kb. The filtration performance was affected by the membrane type used and could be improved by addition of NaCl in the formulation buffer. For filtrations performed at constant pressure, permeate flow decayed with time. As predicted from controlled flux experiments, transmission decreased with increasing molecule size. Initial permeate flux was affected by vector size, DNA concentration and operating pressure. Increase in plasmid size and operating pressure led to reduced membrane capacities. The small scale membrane (filtration area = 1 cm2) capacity was used successfully to predict the performance of a larger scale filtration (filtration area = 4 cm2).

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3

Somridhivej, Benjaporn Liu Zhanjiang. "Characterization, polymorphism assessment, and database construction for microsatellites from BAC end sequences of catfish a resource for integration of linkage and physical maps /." Auburn, Ala., 2007. http://repo.lib.auburn.edu/Send%2011-10-07/SOMRIDHIVEJ_BENJAPORN_30.pdf.

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4

McDowell, Erin. "Characterization of a bacterial artificial chromosome (BAC)-based infectious clone of a low passage Marek's disease virus (MDV) vaccine strain, CVI988." Access to citation, abstract and download form provided by ProQuest Information and Learning Company; downloadable PDF file, 106 p, 2009. http://proquest.umi.com/pqdweb?did=1885544371&sid=6&Fmt=2&clientId=8331&RQT=309&VName=PQD.

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5

Guo, Yufang. "Quantitative genetic analysis for flowering time in primitive Upland cotton, Gossypium hirsutum L., and chromosome assignment of BAC-derived SSR markers." Diss., Mississippi State : Mississippi State University, 2007. http://library.msstate.edu/etd/show.asp?etd=etd-11062007-142438.

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6

Willars, Kate. "Development of targeted bacterial artificial chromosome cloning techniques." Thesis, University of Cambridge, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.620355.

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7

Trapp, Sascha. "Klonierung und Mutagenese des Bovinen Herpesvirus Typ 1 als ein infektiöses künstliches bakterielles Chromosom (bacterial artificial chromosome, BAC)." Diss., lmu, 2003. http://nbn-resolving.de/urn:nbn:de:bvb:19-11529.

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8

Palma, Federica Di. "Analysis and mapping of bovine MHC class I gene." Thesis, University of Reading, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.248145.

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9

Allouis, Sebastien. "Construction of a bacterial artificial chromosome library and gene targeting in pearl millet." Thesis, University of East Anglia, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.327450.

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10

Gill, Clare Alexandra. "Use of an ovine bacterial artificial chromosome library for the study of Bovidae genomes." Title page, table of contents and abstract only, 1999. http://web4.library.adelaide.edu.au/theses/09ANP/09anpg475.pdf.

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11

Wussow, Felix [Verfasser]. "Seamless manipulation of the varicella-zoster virus genome via bacterial artificial chromosome engineering / Felix Wussow." Kiel : Universitätsbibliothek Kiel, 2010. http://d-nb.info/1019951206/34.

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12

Christensen, Heather R. "Molecular and Integrated Systems Physiology of Prolactin." University of Cincinnati / OhioLINK, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1314040076.

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13

Wang, Wenming, Milos Tanurdzic, Meizhong Luo, et al. "Construction of a bacterial artificial chromosome library from the spikemoss Selaginella moellendorffii: a new resource for plant comparative genomics." BioMed Central, 2005. http://hdl.handle.net/10150/610078.

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BACKGROUND:The lycophytes are an ancient lineage of vascular plants that diverged from the seed plant lineage about 400 Myr ago. Although the lycophytes occupy an important phylogenetic position for understanding the evolution of plants and their genomes, no genomic resources exist for this group of plants.RESULTS:Here we describe the construction of a large-insert bacterial artificial chromosome (BAC) library from the lycophyte Selaginella moellendorffii. Based on cell flow cytometry, this species has the smallest genome size among the different lycophytes tested, including Huperzia lucidula, Diphaiastrum digita, Isoetes engelmanii and S. kraussiana. The arrayed BAC library consists of 9126 clones<br>the average insert size is estimated to be 122 kb. Inserts of chloroplast origin account for 2.3% of the clones. The BAC library contains an estimated ten genome-equivalents based on DNA hybridizations using five single-copy and two duplicated S. moellendorffii genes as probes.CONCLUSION:The S. moellenforffii BAC library, the first to be constructed from a lycophyte, will be useful to the scientific community as a resource for comparative plant genomics and evolution.

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14

Taylor, Suzanne. "Towards the construction of a bacterial artificial chromosome (BAC) library of Fragaria vesca L. to isolate a gene controlling seasonal flowering." Thesis, University of Reading, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.369543.

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15

Satorius, Ashley E. "Generation and studies of BKRF4- deficient mutants of Epstein-Barr virus." OpenSIUC, 2010. https://opensiuc.lib.siu.edu/theses/365.

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Epstein-Barr virus (EBV) BKRF4 gene product is a tegument protein encoded by a gene with no sequence homology outside of the gamma subfamily of Herpesviridae. Its positional homologs are necessary for an efficient viral lytic program, in particular viral progeny egress and primary infection. To characterize BKRF4 in this regard, EBV recombinant viruses deficient for BKRF4 were developed using site-directed mutagenesis and a bacterial artificial chromosome (BAC)-based recombineering system. Stable human embryonic kidney (HEK) 293 cell lines containing these genomes were generated and the phenotypes of these mutants were analyzed following stimulation of the viral lytic cycle. During the lytic program, BKRF4-null cell lines showed decreased protein expression of various EBV lytic genes that were analyzed using immunostaining and flow cytometry. Reduced amounts of extracellular viral progeny were observed when quantified by real-time PCR and infectivity assays as compared to wild type. These findings suggest an active role of BKRF4 in EBV infection, possibly in viral egress.

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16

Bambrough, Paul John. "A Bacterial Artificial Chromosome transgenic mouse model allowing assessment of the origin and function of Fibroblast Activation Protein expressing cells in the tumour microenvironment." Thesis, University of Cambridge, 2010. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.608387.

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17

Penha, Helen Alves. "Construção de uma biblioteca genômica de Passiflora edulis f. flavicarpa inserida em BACs (Bacterial Artificial Chromosome) e mapeamento cromossômico usando hibridação in situ fluorescente\"." Universidade de São Paulo, 2012. http://www.teses.usp.br/teses/disponiveis/11/11137/tde-25092012-101518/.

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Passiflora (Passifloraceae) é um grande gênero de espécies vegetais encontradas, principalmente, na flora tropical. Algumas passifloras são cultivadas como plantas ornamentais, frutíferas ou exploradas pelas suas propriedades medicinais. A principal espécie comercial brasileira, o maracujá-azedo (Passiflora edulis f. flavicarpa, 2n = 18), ocupa 95% da área plantada. Os frutos são consumidos in natura ou processados pela indústria de suco. Estudos genéticos e cromossômicos têm sido gerados para esta espécie. Entretanto, devido ao pequeno tamanho e a similaridade morfológica dos seus cromossomos, as medições do cariótipo convencional do maracujá-azedo têm levado a resultados inconsistentes, sendo necessário o desenvolvimento de marcadores cromossomos-específicos. Estes marcadores são produzidos a partir da identificação de sequências de cópia-única em clones de bibliotecas de BACs (Bacterial Artificial Chromosome), que são utilizadas como sondas em ensaios de FISH (Hibridização in situ Fluorescente). Neste trabalho, foi construída uma biblioteca genômica de maracujá-azedo em BACs contendo 82.944 clones, com tamanho médio dos insertos de 108 kb, e provendo uma cobertura de seis vezes o genoma. A biblioteca apresentou baixa contaminação com cpDNA e mtDNA (~0,04% e 0%, respectivamente), e foi possível o isolamento de oito clones contendo genes putativos de P. edulis f. flavicarpa. Estes clones foram marcados e utilizados como sondas em ensaios de FISH. Destas sondas, quatro apresentaram sinal único de hibridização, e foram mapeadas nos cromossomos 1 (gene ERS), 3 (gene ACCO) e 4 (genes G3PD e CYCD1). As demais sondas (genes LOX, NDID e MIPS) apresentaram sinais de hibridização subteloméricos ou pericentroméricos, indicando a presença de DNA repetitivo nos clones; a sonda contendo o gene EMB não revelou sinal fluorescente. Com base em análises de FISH, definiu-se, no presente trabalho, um novo cariótipo para o maracujá-azedo, com marcadores específicos para os cromossomos 1, 3 e 4, localizando, também, sítios de DNAr 45S nos cromossomos 7 e 8, e um sítio de DNAr 5S no cromossomo 5. A exploração da biblioteca de BAC, bem como o mapa físico aqui estabelecido, representa importantes avanços para guiar pesquisas futuras sobre o gênero Passiflora.<br>Passiflora (Passifloraceae) is a large genus of plant species essentially found in the tropical flora. Some passiflora are grown as ornamentals, cultivated for their edible fruits, or exploited due to their medicinal properties. The main Brazilian commercial species, the yellow passion fruit (Passiflora edulis f. flavicarpa, 2n = 18) occupies 95% of all planted orchards. The fruits are eaten fresh or used for industrial juice production. Genetic and chromosomal studies have been carried out on the species. However, due to the small size and morphological similarity of their chromosomes, the conventional measures of the karyotype have produced some inconsistent results, being imperative the development of chromosome-specific markers. These markers are produced by identifying BAC (Bacterial Artificial Chromosome) library clones that harbor single copy sequences, which are used as probes in FISH (Fluorescent in situ Hybridization) assays. In the present work, a yellow passion fruit genomic BAC library of 82.944 clones was constructed, with average insert sizes of 108 kb, and covering six times the genome equivalent. The library has shown a low level of cpDNA and mtDNA contamination (~0.04% and 0%, respectively), and it was possible the isolation of eight clones harboring putative genes of P. edulis f. flavicarpa. These clones were labeled and used as probes in FISH assays. Of these probes, four have shown single hybridization signals, and they were mapped on chromosome 1 (ERS gene), 3 (ACCO gene), and 4 (G3PD and CYCD1 genes). The other probes (LOX, NDID and MIPS gene) revealed subtelomeric or pericentromeric signals, suggesting the presence of repetitive DNA sequences in the clones; the probe harboring the EMB gene did not reveal any hybridization signal. Based on FISH analyses, a new karyotype for the passion fruit was established in the present work, with specific markers in chromosomes 1, 3 and 4; we also mapped 45S rDNA sites in chromosomes 7 and 8, and one 5S rDNA site in chromosome 5. The exploitation of the BAC library, as well as the physical map here established, represents novel and essential advances to guide future researches on the Passilfora genus.

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18

Xu, Zhiyong [Verfasser]. "Generation of a Complete Single-Gene Knockout Bacterial Artificial Chromosome Library of Cowpox Virus and Identification of Genes involved in hemorrhagic lesions on infected chicken chorioallantoic membranes (CAMs) / Zhiyong Xu." Berlin : Freie Universität Berlin, 2015. http://d-nb.info/106486953X/34.

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19

Bittorf, Blaine E. "Mapping Hybrid Lethal Genes on the X Chromosome of C. Briggsae." Wright State University / OhioLINK, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=wright152770556182685.

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20

Dougherty, John Kelly. "Identification of a Hybrid Lethal Gene on the X Chromosome of Caenorhabditis briggsae." Wright State University / OhioLINK, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=wright1579011194671611.

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21

Thomas, Matthew Robert. "Functional analysis of the ALS/FTD associated gene FUS using a novel in vitro genomic DNA expression system." Thesis, University of Oxford, 2013. http://ora.ox.ac.uk/objects/uuid:176cbd1b-623e-41cb-9237-cd6c82fc0ad4.

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Aggregations of fused in sarcoma (FUS), a multifunctional RNA processing protein, define a pathological subtype of both frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS), whilst mutations in the FUS gene are causative for ALS. To model the impact of FUS mutations, expression vectors containing the entire genomic sequence of FUS, up and downstream regions, and native promoter sequences have been generated. The constructs have been tagged with an mCherry fluorescent tag, and three separate pathological mutations (R244C, R521C, and P525L) have been separately inserted. Transgenic mice have been generated using the WT and P525L FUS vectors to provide a highly physiological model of FUS in disease. Within transfected HEK293 cells, insertion of the P525L and R521C FUS mutations leads to relocalisation of FUS from the nucleus to the cytoplasm. R521C and P525L mutant FUS incorporates into cytoplasmic aggregations of untranslated mRNA and RNA binding proteins known as stress granules. The strong relocalisation seen with P525L-FUS is associated with a gain of cytotoxicity. Reversal of this cytoplasmic relocalisation by demethylation of FUS rescues this cytotoxicity, suggesting a toxic gain of cytoplasmic function in the majority of FUS mutations. By contrast, insertion of the R244C mutation leads to neither relocalisation, stress granule association, nor cytotoxicity. Notably the R244C mutation, located away from the nuclear localization domain in which the majority of FUS mutations are found, leads to the presence of smaller FUS fragments in western blot analyses. These fragments appear not to be due to splicing defects in FUS but rather are due to post-translational modifications or aberrant protein cleavage. These data suggest an alternative pathway for FUS toxicity based upon a nuclear loss of function.

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22

Mammadov, Jafar. "Towards Cloning the Leaf Rust Resistance Gene Rph5." Diss., Virginia Tech, 2004. http://hdl.handle.net/10919/28704.

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Leaf rust caused by Puccinia hordei is an important disease of barley (Hordeum vulgare) in many regions of the world. Yield losses up to 62% have been reported in susceptible cultivars. The Rph5 gene confers resistance to the most prevalent races (8 and 30) of barley leaf rust in the United States. Therefore, the molecular mapping of Rph5 is of great interest. Genetic studies were performed by analysis of 93 and 91 F2 plants derived from the crosses 'Bowman' (rph5) x 'Magnif 102' (Rph5) and 'Moore' (rph5) x Virginia 92-42-46 (Rph5), respectively. Linkage analysis positioned the Rph5 locus to the extreme telomeric region of the short arm of barley chromosome 3H at 0.2 cM proximal to RFLP marker VT1 and 0.5 cM distal from RFLP marker C970 in the Bowman x Magnif 102 population. Synteny between rice chromosome 1 and barley chromosome 3 was employed to saturate the region within the sub-centimorgan region around Rph5 using sequence-tagged site (STS) markers that were developed based on barley expressed sequence tags (ESTs) syntenic to the phage (P1)-derived artificial chromosome (PAC) clones comprising distal region of the rice chromosome 1S. Five rice PAC clones were used as queries to blastn 370,258 barley ESTs. Ninety four non-redundant EST sequences were identified from the EST database and used as templates to design 174 pairs of primer combinations. As a result, 10 EST-based STS markers were incorporated into the 'Bowman' x 'Magnif 102' high-resolution map of the Rph5 region. More importantly, six markers, including five EST-derived STS sequences, co-segregate with Rph5. Genes, represented by these markers, are putative candidates for Rph5. Results of this study demonstrate the usefulness of rice genomic resources for efficient deployment of barley EST resources for marker saturation of targeted barley genomic region.<br>Ph. D.

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23

DALÍKOVÁ, Martina. "Využití BAC klonů při studiu pohlavního chromosomu W obaleče jablečného \kur{Cydia pomonella} (Lepidoptera: Tortricidae)." Master's thesis, 2009. http://www.nusl.cz/ntk/nusl-49849.

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In the present study, the W sex-chromosome of the codling moth was studied by means of fluorescence in situ hybridization (FISH) with probes prepared from bacterial artificial chromosome (BAC), which were isolated from the codling moth BAC library. The BAC library was screened for clones derived from both the W and Z sex chromosomes using three sets of molecular markers of codling moth sex chromosomes. A total of 54 BAC clones have been obtained. In this work, only 3 W-derived BAC clones and 1 Z-derived BAC clone were further characterized by BAC-FISH mapping on chromosome preparations of pachytene oocytes; the other BAC clones have been retained for next studies. Whereas the Z-BAC probe provided a discrete hybridization signal on the Z chromosome, and surprisingly on the W chromosome, the W-BAC probes showed multiple hybridization signals distributed on the whole W chromosome, suggesting that they are mainly composed of repetitive sequences, which occur in multiple clusters on the W chromosome. The specific pattern of W-BAC hybridization signals along with the discrete signal of the Z-BAC enabled us to discriminate left/right orientation of both the W and Z chromosomes and examine specificity of W-Z pairing during meiotic prophase I.

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24

Vichai, Vanicha. "Mouse class-mu glutathione transferase gene expression, structure and organization /." 2000. http://wwwlib.umi.com/dissertations/fullcit/9975515.

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25

Limaye, Nisha. "Common alleles of the SLAM/CD2 family are associated with murine lupus." 2005. http://edissertations.library.swmed.edu/pdf/LimayeN042905/LimayeNisha.pdf.

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26

Nikiforuk, Aidan. "Generation of a human Middle East respiratory syndrome coronavirus (HCoV-MERS) infectious clone system by recombination of bacterial artificial chromosomes." 2015. http://hdl.handle.net/1993/30635.

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Coronaviruses have caused high pathogenic epidemics within the human population on two occasions; in 2003 a coronavirus (HCoV-SARS) caused severe acute respiratory syndrome and in 2012 a novel coronavirus emerged named Middle East respiratory syndrome (HCoV-MERS). Four other species of coronavirus circulate endemically in the human population (HCoV-229E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1), which cause more benign respiratory disease than either HCoV-SARS or HCoV-MERS. The emergence of HCoV-MERS provides an additional opportunity to study the characteristics of coronaviruses. Reverse genetics can be used to study an organism’s phenotype by logical mutation of its genotype. Construction of an infectious clone construct provides a means to investigate the nature of HCoV-MERS by reverse genetics. An HCoV-MERS infectious cDNA clone system was constructed to use for reverse genetics by homologous recombination of bacterial artificial chromosomes (BACs). This system should aid in answering remaining questions of coronavirus genetics and evolution as well as expedite the development of vaccines and prophylactic treatments for HCoV-MERS.<br>October 2015

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27

Bachmann, J. A., Andrew Tedder, B. Laenen, K. A. Steige, and T. Slotte. "Targeted long-read sequencing of a locus under long-term balancing selection in Capsella." 2018. http://hdl.handle.net/10454/17277.

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Yes<br>Rapid advances in short-read DNA sequencing technologies have revolutionized population genomic studies, but there are genomic regions where this technology reaches its limits. Limitations mostly arise due to the difficulties in assembly or alignment to genomic regions of high sequence divergence and high repeat content, which are typical characteristics for loci under strong long-term balancing selection. Studying genetic diversity at such loci therefore remains challenging. Here, we investigate the feasibility and error rates associated with targeted long-read sequencing of a locus under balancing selection. For this purpose, we generated bacterial artificial chromosomes (BACs) containing the Brassicaceae S-locus, a region under strong negative frequency-dependent selection which has previously proven difficult to assemble in its entirety using short reads. We sequence S-locus BACs with single-molecule long-read sequencing technology and conduct de novo assembly of these S-locus haplotypes. By comparing repeated assemblies resulting from independent long-read sequencing runs on the same BAC clone we do not detect any structural errors, suggesting that reliable assemblies are generated, but we estimate an indel error rate of 5.7×10−5. A similar error rate was estimated based on comparison of Illumina short-read sequences and BAC assemblies. Our results show that, until de novo assembly of multiple individuals using long-read sequencing becomes feasible, targeted long-read sequencing of loci under balancing selection is a viable option with low error rates for single nucleotide polymorphisms or structural variation. We further find that short-read sequencing is a valuable complement, allowing correction of the relatively high rate of indel errors that result from this approach.<br>This study was supported by a grant from the Swedish Research Council to T.S.

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28

Anderson, Jason C. "Cytogenomic Analyses of the genus Sorghum." Thesis, 2010. http://hdl.handle.net/1969.1/ETD-TAMU-2010-05-7847.

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A phylogenetic tree based on ITS1, Adh1 and ndhF grouped the species of the genus Sorghum into one distinct monophyletic group, but including two sister lineages, one with x=5, the other with x=10 as basic chromosome numbers. The goal of this study was to elucidate major patterns in Sorghum genome evolution, particularly n=5 vs. n=10 genomes. A very recent molecular cytogenetic study in our laboratory revealed striking structural karyotypic rearrangements between S. bicolor (x=10) and an x=5 Sorghum species, S. angustum; so an immediate objective here was to determine if identical or similar rearrangements exist in other wild Sorghum species. Our approach was [1] to extend similar methods to additional species, i.e., fluorescent in situ hybridization (FISH) analyses of sorghum genomic bacterial artificial chromosome clones and multi-BAC cocktail probes to mitotic chromosomes of S. angustum, S. versicolor, S. brachypodum and S. intrans; and [2] to augment the BAC-FISH findings by comparing telomeric and ribosomal DNA FISH signal distributions to x=5 and x=10 Sorghum species. Signals from in situ hybridizations of BAC-based probes were insufficiently robust and insufficiently localized to delineate FISH signal patterns akin to those discovered previously in S. angustum. Southern blots of the same BACs to restricted DNA of these species revealed relatively moderate affinity to smeared DNA, suggesting homology to non-tandemized sequences. FISH of the A-type TRS (Arabidopsis-like telomeric repeat sequence) revealed its presence is limited to terminal chromosomal regions of the Sorghum species tested, except S. brachypodum, which displayed intercalary signal on one chromosome and no detachable signal at its termini region. The hybridization of 45S and 5S rDNA revealed that the respective sites of tandemized clusters differ among species in terms of size, number and location, except S. angustum versus S. versicolor. Well localized BAC-FISH signals normally occur when signals from low-copy sequences discernibly exceed background signal, including those from hybridization of dispersed repetitive elements. The low level of signal intensity from BAC low-copy sequences relative to the background signal "noise" seems most likely due to low homology and(or) technical constraints. Extensive dispersal of low-copy sequences that are syntenic in S. bicolor seems unlikely, but possible. In conclusion, the result was a lack of clear experimental success with BAC-FISH and an inability to effectively screen for S. angustum-like rearrangements using BAC-FISH. The telomeric and rDNA FISH indicated that the x=5 genomes vary extensively. One can surmise that although the arrangements seen in S. angustum might extend to S. versicolor, they certainly do not extend to S. versicolor, they certainly do not extend to S. intrans or S. brachypodum. It is clear that S. brachypodum has telomeric repeats that are either very short or rely on some sequence other than the A-type TRS.

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29

Trapp, Sascha [Verfasser]. "Klonierung und Mutagenese des bovinen Herpesvirus Typ 1 als ein infektiöses künstliches bakterielles Chromosom (bacterial artificial chromosome, BAC) / von Sascha Trapp." 2003. http://d-nb.info/968529682/34.

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30

Wu, Pei-Yu, and 吳沛宇. "Construction of the full-length pseudorabies virus TNL strain genome as the bacterial artificial chromosome." Thesis, 2002. http://ndltd.ncl.edu.tw/handle/59281304081566925981.

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碩士<br>國立中興大學<br>獸醫學系<br>90<br>Construction of full-length infectious clones of virus genome is useful for the study of viral pathogenesis and the application of vaccine development. We used the recently developed bacterial artificial chromosome (BAC) as a vector to carry 150 kb full-length genome of the pseudorabies TNL strain (a Taiwan local strain). A green fluorescent protein (eGFP) expression cassette flanked by two 34 base loxP sites was inserted into the segment of 11 k and 28 k genes for the homologous recombination. After 15 passages, green plaques were cloned and named TNL-GFP. By using the method of cre-loxP site-specific recombination, dark plaques, the TNL-loxP, were picked and proved to have the loxP site insertion of PRV genome by sequencing. We modified pCC1TM BAC vector from Epicentre to be inserted with another GFP cassette. This BAC vector containing a loxP site and the GFP expression cassette was integrated into PRV genome again by cre-loxP recombination thus resulted in green fluorescent plaques, named PRV-BAC. By plaque assay, we found the insertion of a GFP cassette between 11 k and 28 k genes would reduce the viral titer. Moreover, another large insertion as BAC vector could decrease the plaque size of PRV。By sequencing of the PCR fragment, we sugessted there might be rearrangements in TNL-BAC genome, thus the inserted GFP fragment could not be detected by PCR. Therefor the insertion of BAC vector was still identified by the dot-blot hybridization with linearised pCC1 as probe。After transforming E. coli with TNL-BAC infected cell DNA by electroporation, the plasmids from chloramphenicol resistant colonies were purified. However, these plasmids can only be detected to have the pCC1 sequence by dot-blot hybridization, but they didn’t show any detectable PRV-TNL sequence by PCR.

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31

Schäfer, Martin [Verfasser]. "Human DC-SIGN mediated recognition of mycobacteria in conventional and bacterial artificial chromosome transgenic mouse models / Martin Schäfer." 2006. http://d-nb.info/985177160/34.

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32

Nagel, Claus-Henning [Verfasser]. "Fluorescence tagging of herpes simplex virus type 1 proteins by mutagenesis of a bacterial artificial chromosome / von Claus-Henning Nagel." 2006. http://d-nb.info/98195233X/34.

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Özyurt, Baris. "Identifikation von Genen und Mikroorganismen, die an der dissimilatorischen Fe(III)-Reduktion beteiligt sind." Doctoral thesis, 2009. http://hdl.handle.net/11858/00-1735-0000-0006-B66A-8.

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