Counter selectable allelic exchange
WebFeb 27, 2012 · In a typical allelic exchange experiment, the chromosomal sequence to be mutated is synthesized and cloned onto a vector. This sequence is either highly … WebApr 7, 2024 · SacB gene encoding a levansucrase, which can convert sucrose into a toxic metabolite, is the most commonly used counter-selectable marker in C. glutamicum. Schäfer et al. (1994) successfully …
Counter selectable allelic exchange
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WebNov 1, 2004 · The adaptation of the counter-selectable marker pheS*, coding for a mutated subunit of the phenylalanine tRNA synthetase, for use in S. aureus, and the new allelic exchange plasmid pIMAY*, which will aid the efficient and easy creation of gene knockouts. 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 …
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 … 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
Weban 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 … WebFeb 16, 2016 · We believe that a suicide plasmid should have another visual biomarker as a counter selectable tool. Thus, in this study, to improve the efficiency of allelic exchange for genome genetic modification, a novel …
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 …
WebSep 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 … cp bella vista corrientesWebAug 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). … cp belcantoWebJan 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 … cp bellpratWebDec 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 … cp bella vista buenos airesWebNov 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 ... cpb essential 作用WebOne key methodology is the allelic exchange of unmarked, in-frame deletion mutations and point mutations using counter-selectable suicide plasmids. I describe below the challenges of allelic exchange in M. tuberculosis, the overall mechanism of a suicide vector system, and how it can be used for efficient allelic exchange in this formally ... cp bell-llocWebOct 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... cpbf montagnac