Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 16 of 16 for “"recombineering"”.
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A recombineering pipeline for functional genomics applied to Caenorhabditis elegans
… authentic regulation with a new application of recombineering, which is simple, fast and efficient. For the first time we apply liquid culture cloning for multiple recombineering steps. This is particularly important when high throughput applications are considered, as it offers significant …
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Adding to the recombineering toolbox: interrogating internal transcriptional complexity in Caenorhabditis elegans operons via recombineered fosmid-based reporters
… reporters via seamless counter-selection recombineering two resource sets were first constructed. The first of these contained FP coding sequences synthesized de novo and codon-optimized for expression in C. elegans. The second set comprised a series of constructs designed to streamline …
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Development of Improved Genetic Methods for Enterococcus Faecium
… significantly improves the efficiency of recombineering technologies in both commensal and antibiotic-resistant strains of E. faecium and other Enterococcus species such as E. durans and E. hirae. Here, I utilize a recombineering system using a latent bacterial prophage RecT that is …
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Development of embryonic stem cells expressing endogenous levels of a fluorescent protein fused to the telomere binding protein TRF1
… Trf1 promoter. To do this, I have used a recombineering technique using Bacterial Artificial Chromosomes (BACs). BAC recombineering allows for the direct knock-in of a fluorescent tag into the mouse Trf1gene locus. Genetic constructs with the correct sequence inserts have been obtained and …
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BAC transgene arrays as a model system for studying large-scale chromatin structure
… targets to the nuclear periphery. Using BAC recombineering, we dissected this 200kb region and identified two genomic regions sufficient to target the BAC transgenes to nuclear periphery. This study represents a first step towards elucidation of the molecular mechanism for the nuclear …
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Algorithms for E. coli genome engineering
Author summary: Lamba red recombineering is one of methods of performing genome engineering. However, this method of genome editing is not very specific and efficient and is highly dependent on the genomic regions that are targeted (integration sites). In this project we explored ways of …
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Construction of a COL11A1 Transgene Vector
… Results: Through the relatively new approach of "recombineering", the coding sequence of a human COL11A1 cDNA was constructed from two cloned cDNA fragments. A copy of the cDNA was inserted into the pcDNA3.1 expression vector for in vitro transcription/translation assays. Another copy of the cDNA …
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Developing High Purity Embryonic Stem Cell-Derived V2a Interneurons for In Vitro Investigation and Transplantation Following Spinal Cord Injury
… Using CRISPR/Cas9-mediated homologous recombineering, we generated a transgenic mouse embryonic stem cell (ESC) line that enabled us to positively select for Chx10+ ESC-derived V2a interneurons (ESC-V2as) by treating differentiated cells with puromycin. ESC-V2a cultures remained free of …
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Generation of genetically modified mice with conditional alleles of Dyrk1a : a minimal animal model of Down syndrome
… nutzten. Die neue Technik des „Recombineering“ ermöglichte eine schnelle und effiziente Herstellung von Mäusen mit einem loxP flankierten Dyrk1a Allel. Die heterozygoten Dyrk1a(flox/+) Mäuse und die homozygoten Dyrk1a(flox/flox) Mäuse sind lebensfähig und sie zeigen bisher keine …
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Generation and Characterization of a Knock-In Allele of EKLF: Probing the in vivo Role of the Chromatin Remodeling Domain in Definitive Hematopoietic Cells
… a knock-in allele of D221EKLF. Using the recombineering method, a lambda phage-based homolgous recombination method in <em>E. coli,</em> cDNA encoding theD221EKLF domain has been inserted into the endogenous initiation site, thus placing the mutant protein under the <em>cis-</em>regulatory …
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Functional genomic analysis of cell cycle progression in human tissue culture cells
… normal gene expression. In combination with recombineering strategies this technology is useful for RNAi rescue experiments, protein localisation and the identification of protein interaction partners in mammalian tissue culture cells. In summary, this thesis presents a major technical …
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Chemical genetic manipulation of interferon regulatory factor 1 (IRF-1) using synthetic biology
… Firstly, we described the creation, using recombineering technology, of universal bacterial artificial chromosome (BAC) targeting vector. This targeting vector, carry a cre-conditioned STOP cassette that can be targeted at a desired specific area. The resulted targeting vector can aid the …
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The immunological role of cell wall components from diverse Mycobacterium tuberculosis clinical isolates in regulating HIV-1 replication in human macrophages
… subject to single nucleotide mutagenesis using recombineering to either revert SNP of interest to match the alleles of H37Rv or introduce the SNP of interest into the control strains. The wild-type and mutant strains were used in a trans-well assay to infect MDM in the presence of HIV-1 …
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Engineering Zymomonas mobilis for the production of biofuels and other value-added products
Zymomonas mobilis is a promising organism for lignocellulosic biofuel production as it can efficiently produce ethanol from simple sugars using unique metabolic pathways. Z. mobilis displays what is known as the “uncoupled growth” phenomenon, meaning cells will rapidly convert sugars to ethanol …
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Defining the Requirements for Early Gene Expression in Bacteriophage HK639
<p>Lambdoid phages suppress transcription termination to fully express their genes. Antitermination of early gene expression in most lambdoid phages is mediated by an interaction between the N protein and a number of host-encoded factors. Bacteriophage HK022 does not rely on a protein for …
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Investigating the Role of Human Cytomegalovirus Protein LUNA in Regulating Viral Gene Expression during Latency
Human cytomegalovirus (HCMV) is a widespread human herpesvirus pathogen and prototypical member of the β-herpesvirus subfamily. Like all herpesviruses, the virus establishes a lifelong latent infection following host exposure, which has the potential to reactivate periodically and contribute to …