Counter selectable allelic exchange

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August undefined, 2024

WebOct 25, 2024 · Counter-selectable markers are genes that promote the death or severe growth inhibition of host cells under appropriate growth conditions [ 1 ]. These markers are useful genetic tools for a variety of applications, including creation of chromosomal mutations [ 2, 3, 4, 5 ], selection of recombinant vectors [ 6, 7 ], and plasmid curing [ 8, 9, … WebNov 1, 1996 · With this methodology, a sucrose counter-selectable vector was used to deliver, into the Mycobacterium bovis BCG genome, an inactivated copy (ureC::Km) of …

Allelic exchange of unmarked mutations in Mycobacterium

WebNov 1, 1996 · Re- cently, two counter-selectable markers have been de- scribed in M. smegmatis and used to demonstrate that positive selection of allelic exchange mutants is possible m this bacterium: (i) the rpsL gene confer- ring a dominant phenotype of sensitivity to strepto- mycin [6], and (ii) the Bacillus subtilis sacB gene con- ferring sensitivity to ... WebNov 1, 1996 · With this methodology, a sucrose counter-selectable vector was used to deliver, into the Mycobacterium bovis BCG genome, an inactivated copy (ureC::Km) of the ureC gene encoding the mycobacterial urease. A two-step selection procedure on 2% sucrose allowed the positive selection of gene exchange mutants. picture of rugby pitch

Positive selection of allelic exchange mutants in ... - PubMed

WebThis chapter describes the method of allelic exchange in Staphylococcus aureus using temperature-sensitive plasmids that have successfully produced a variety of … WebJan 1, 2011 · To make use of rps L gene as a counter-selectable marker system instead of sac B for efficient selection of allelic exchange mutants in C. glutamicum, both a host strain YHIK-18, which corresponds to the rps L + mutant of C. glutamicum ATCC13032, and a new non-replicative integration vector pCGI harboring rps L - gene under control of the … WebNov 5, 2024 · Existing methods for genetic manipulation of A. baumannii mainly rely on the use of antibiotic as the selectable marker, and the sacB /sucrose as the counter-selectable marker, which is ... picture of rupaul\u0027s wife

Harnessing Bacterial Toxins for Allelic Exchange

Knock-out or knock-in? Converting a SacB-based gene disruption s…

Weballele of interest (a or b). For excision to lead to a successful allelic exchange, recombinalion must occur in the second region of homology. Note that strains thai have … WebHarnessing Bacterial Toxins for Allelic Exchange. Type. ... Modifying bacterial genomes using counter-selectable markers and homologous recombination As an alternative... picture of rudolph in real lifeWebAn allelic exchange protocol based on double crossover recombination facilitated by the Bacillus subtilis sacB gene that can be applied for partial or complete gene knockouts, … picture of running shoes

"WebOct 25, 2024 · A protocol was designed for plasmid curing using a novel counter-selectable marker, named pylSZK-pylT, in Escherichia coli. The pylSZK-pylT marker … " - Counter selectable allelic exchange

Counter selectable allelic exchange

Development of an efficient technique for gene deletion and allelic ...

WebFeb 28, 2014 · In this study, a gfp-based novel markerless allelic exchange integration system was developed.By employing gfp gene and sacB gene as counter-selectable … WebJan 1, 2006 · To test whether pKOR1 permits allelic replacement without antibiotic marker selection in S. aureus, in-frame deletions were generated in the rocA gene of the human …

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WebOct 22, 2015 · Allelic exchange is an efficient method of bacterial genome engineering. This protocol describes the use of this technique to make gene knockouts and knock-ins, as well as single-nucleotide... WebDec 1, 2024 · Counter-selectable markers are genes that promote the death or severe growth inhibition of host cells under appropriate growth conditions [1]. ... To facilitate …

WebJul 17, 2024 · This is the first step of the two-step allelic exchange. A second homologous recombination event removes the integrated vector … Web(Empty Backbone) Allelic exchange plasmid with PheS* counter-selection for Staphylococcus aureus Depositing Lab. Angelika Grundling. Publication. Schuster et al Microbiology. 2024 Apr 3. doi: 10.1099/mic.0.000791. ... Use of the counter selectable marker PheS* for genome engineering in Staphylococcus aureus.

WebAug 15, 2024 · Bacterial toxins for counter-selection. In 2015, the Chen lab harnessed a group of bacterial toxins to facilitate efficient recombineering (Khetrapal et al., 2015). … Website-specific mutagenesis by allelic exchange was evaluated. To develop this system, we optimized conjugal DNA transfer by isolating a mutant that functions 1000-fold more efficiently as a recipient than the wild type strain, identified two counter- selectable markers, rpsL and sacB, that function

WebApr 5, 2024 · Counter-selection methods have been developed for thermophilic organisms to select for the second homologous recombination event and hence reduce the screening process. One such method is the pyrF based system [14, 15]. This technique utilises the toxicity to 5-fluoroorotic acid conveyed by the pyrF gene product to select for integration …

WebApr 10, 2024 · The Golden Gate method and the counter selectable marker were combined innovatively. Abstract. To investigate gene function in Bacillus anthracis, a high-efficiency cloning system is required with an increased rate of allelic exchange. Golden Gate cloning is a molecular cloning strategy allowing researchers to simultaneously and … top gear amazon prime removed 2022WebApr 3, 2024 · Here, a host-genotype-independent counter-selectable marker pheS* was constructed through a point mutation of the gene pheS, which encodes the α-subunit of … picture of rudolph the red-nosed reindeerWeban efficient counter-selector during gene knockout proce-dures in S. aureus. In this study, we synthetically adapted the pheS* gene for counterselection use in S. aureus and … picture of running horsesWebJul 1, 2024 · Pseudomonas aeruginosa is a key opportunistic human pathogen. An established procedure to replace a target gene is two-step allelic exchange, i.e. selection of single crossover at homologous sequences and subsequent counter selection to induce double crossover for excision of the suicide vector. In this study, we found that certain … picture of ruby stoneWebSep 19, 2024 · Herein we describe an optimized and rapid method to create unmarked deletion mutants of P. aeruginosa in as few as seven days with total hands-on time of less than half a day. Briefly, allelic exchange constructs for gene deletion are generated by Gibson assembly [ 9] and introduced into P. aeruginosa using an optimized … picture of rupp arenaWebJan 7, 2024 · In the last 10 years, the barriers preventing the uptake of foreign DNA by clinical Staphylococcus aureus isolates have been identified and powerful mutagenesis … picture of rush doors robloxWebOct 1, 2002 · The authors have developed a simple and highly efficient system for generating allelic exchanges in both fast- and slow-growing mycobacteria. In this procedure a gene of interest, disrupted by a selectable marker, is cloned into a conditionally replicating (temperature-sensitive) shuttle phasmid to generate a specialized transducing … picture of rural area