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Che9c

WebPhage Che9c gp61-mediated recombination is sufficiently efficient that single base changes can be introduced without requirement for direct selection, with isogenic mutant strains … WebBiochemical characterization of Che9c gp60 and gp61 demonstrated that they possess exonuclease and DNA binding activities, respectively, similar to RecET and λ Exo/Beta. …

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WebChe9c gp61 is clearly a member of the superfamily of SSAPs 30 with 29% amino acid identity to E. coli RecT, but with a longer C-terminal extension (Fig. 1a). Although Che9c … WebG@ Bð% Áÿ ÿ ü€ H FFmpeg Service01w ... e-way portal login https://seelyeco.com

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WebExpression and biochemical characterization show that Che9c gp60 and gp61 encode exonuclease and DNA-binding activities, respectively, and expression of these proteins substantially elevates recombination facilitating allelic exchange in both M. smegmatis and M. tuberculosis. Mycobacterial recombineering thus provides a simple approach for the ... WebJan 16, 2024 · Che9c RecT annealase from the anhydrotetracycline (ATc)-inducible Ptet promoter (pKM402) 39. The frequency of oligo incorporation was determined as the number of HygR transformants among the survivors of electroporation. The number of HygR transformants generated varied from less than 10 to over 300, depending on the amount … WebAug 17, 2024 · Using Che9c_60 as a model for RecE-like exonuclease (from approved list), HHPred reveals matches to 3H4R_A and PF12705.7. Che9c_61 is called as a RecT-like ssDNA binding protein. There are no alignments to anything labeled as RecE. eway platform

Reprogramming the endogenous type III-A CRISPR-Cas …

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Che9c

Mycobacterial Recombineering Springer Nature Experiments

WebBiochemical characterization of Che9c gp60 and gp61 demonstrated that they possess exonuclease and DNA binding activities, respectively, similar to RecET and λ Exo/Beta. Expression of gp60/gp61 in M. smegmatis and M. tuberculosis substantially increases homologous recombination such that 90% of recovered colonies are the desired gene ... WebMar 14, 2016 · The helper plasmid pJV53, which expresses the Che9c genes, is widely used for gene recombination 8. Following recombination, cells transformed with pJV53 are plated on sucrose-containing media to ...

Che9c

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WebChe9c gene 41 encodes a tyrosine-integrase and a putative Xis encoded by gene 50, displaced almost 7 kbp from the int gene. A putative attP common core can be identified … WebJan 1, 2015 · A second protocol described here is the methodology of using the Che9c RecT function in transferring single base pair mutations, additions or deletions to the M. …

WebChe9c gp60 has exonuclease activity in vitro that is dependent on the presence of dsDNA ends, and Che9c gp61 binds short (20 nucleotides) … WebWhen both Che9c gp60 and gp61 are expressed from an inducible expression system in M. smegmatis or M. tuberculosis, recombination frequencies are elevated substantially. Introduction of a dsDNA allelic exchange substrate in which 500–1000 bp of chromosomal homology flank an antibiotic resistance marker, ...

http://tdelia-irsc.weebly.com/uploads/1/2/1/2/12124315/function_nomenclature_2016.pdf Web(A) The genome maps of phages Che9c and Sbash are shown with shared nucleotide sequence similarity represented as spectrum-colored shading between the genomes, with violet representing the most ...

WebThe Che9c phage also encodes a function similar to RecT. Specifically, the C-terminal domain of Che9c gp60 is 28% identical to a nuclease domain in the RecB family of …

WebExpression and biochemical characterization show that Che9c gp60 and gp61 encode exonuclease and DNA-binding activities, respectively, and expression of these proteins … eway portal gov inWebJul 7, 2024 · Che9c is the only known phage to which Charlie gp32 provides defense. In a second example, phage Phrann codes for a putative (p)ppGpp synthetase (gp29) and gp30 which when expressed together from a prophage confer defense against several unrelated phages including phage Tweety . Expression of Phrann gp29/gp30 strongly inhibits … bruce tinkoff barrington ilWebAll dsDNA phages encode two proteins involved in host lysis, an endolysin and a holin that target the peptidoglycan and cytoplasmic membrane, respectively. Bacteriophages that … eway pin code distanceWebPlasmid pJV53 from Dr. Graham Hatfull's lab contains the insert Che9c genes 60-61 and is published in Nat Methods. 2007 Feb . 4(2):147-52. This plasmid is available through … bruce tinkoff attorneyWebThis chapter describes a protocol for the generation of defined mutants using recombination tools in an inducible recombination system driven by mycobacterial Che9c phage RecET proteins, originally developed in Dr. Graham Hatfull's group, combined with linearized recombination substrates containing flanking sequences of a locus of interest and ... eway portal downWeb8 Likes, 1 Comments - Modnaya_odejda (@tashkent_modnayaodejda) on Instagram e way portal gov inWebApr 1, 2024 · Bacteriophage Che9c gp60 has dsDNA exonuclease activity, degrading linear dsDNA templates with free ends, and Che9c gp61 binds to short (50-base) ssDNA substrates [7 ••, 17, 18]. Briefly, electrocompetent cells expressing the recombination proteins (i.e. gp60 and gp61) are co-electroporated with the recombination substrate and … bruce tinsley