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The pBAD/His Kit provides all of the necessary reagents to express your protein in a tightly regulated fashion. The vector pBAD/His allows you to express your protein with an N-terminal tag. The vector provides:The araBAD pro

Abstract. In vivo induction of the Escherichia coli lactose operon as a function of inducer concentration generates a sigmoidal curve, indicating a non-linear r

Nature Genetics - A new logic for DNA engineering using recombination in

We report the identification and functional analysis of nine genes from Gram-positive and Gram-negative bacteria and their phages that are similar to lambda (λ) bet or Escherichia coli recT . Beta and RecT are single-strand DNA annealing proteins, referred to here as recombinases. Each of the nine other genes when expressed in E. coli carries out oligonucleotide-mediated recombination. To our knowledge, this is the first study showing single-strand recombinase activity from diverse bacteria. Similar to bet and recT , most of these other recombinases were found to be associated with putative exonuclease genes. Beta and RecT in conjunction with their cognate exonucleases carry out recombination of linear double-strand DNA. Among four of these foreign recombinase/exonuclease pairs tested for recombination with double-strand DNA, three had activity, albeit barely detectable. Thus, although these recombinases can function in E. coli to catalyze oligonucleotide recombination, the double-strand DNA recombination activities with their exonuclease partners were inefficient. This study also demonstrated that Gam, by inhibiting host RecBCD nuclease activity, helps to improve the efficiency of λ Red-mediated recombination with linear double-strand DNA, but Gam is not absolutely essential. Thus, in other bacterial species where Gam analogs have not been identified, double-strand DNA recombination may still work in the absence of a Gam-like function. We anticipate that at least some of the recombineering systems studied here will potentiate oligonucleotide and double-strand DNA-mediated recombineering in their native or related bacteria.

Homologous DNA recombination is a fundamental, regenerative process within living organisms. However, in most organisms, homologous recombination is a rare event, requiring a complex set of reactions and extensive homology. We demonstrate in this paper that Beta protein of phage λ generates recombinants in chromosomal DNA by using synthetic single-stranded DNAs (ssDNA) as short as 30 bases long. This ssDNA recombination can be used to mutagenize or repair the chromosome with efficiencies that generate up to 6% recombinants among treated cells. Mechanistically, it appears that Beta protein, a Rad52-like protein, binds and anneals the ssDNA donor to a complementary single-strand near the DNA replication fork to generate the recombinant. This type of homologous recombination with ssDNA provides new avenues for studying and modifying genomes ranging from bacterial pathogens to eukaryotes. Beta protein and ssDNA may prove generally applicable for repairing DNA in many organisms. * oligo(s), : oligonucleotide(s); dsDNA, : double-stranded DNA; ssDNA, : single-stranded DNA

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WIZ AI Talkbot Software-as-a-Service (SaaS) Terms and Conditions THIS SOFTWARE AS A SERVICE AGREEMENT (“THE “AGREEMENT”) IS ENTERED INTO BETWEEN CUSTOMER AND WIZ. CUSTOMER AND WIZ AGREE THAT THE FOLLOWING TERMS AND CONDITIONS WILL APPLY TO THE SERVICES PROVIDED UNDER THIS AGREEMENT.1. Definition and Interpretation1.1 In this Agreement, the following words and expressions shall have […]

Chromosome structure is a key regulator of gene expression. CTCF and cohesin play critical roles in structuring chromosomes by mediating physical inte…

Cloud security startup Wiz has raised more than $120 million in a funding round led by customer relationship management (CRM) giant Salesforce, according to Bloomberg.

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