NATX Plasmids, for: DNA Vaccines, Gene Therapeutics, Viral Vectors andBiophamacueticals

NTC Recombineering Vectors: Chromosome Engineering in E. coli

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Versions
pKD46-recA
pKD46-recApa
XXDescription


Recombineering (recombinogenic engineering) is a homologous recombination-based technology used to modify DNA. Target DNA molecules (plasmids, BAC vectors or the host chromosome) are precisely altered by homologous recombination in host cells which express recombineering enzymes. Recombineering in E. coli often utilize the phage λ Red recombination functions (Murphy, 1998; Datsenko and Wanner, 2000).  The λ genes involved in Red recombination are exo, bet, and gam. The exo (Reda) gene product has 5′ to 3′ exonuclease activity, and the bet (Redb) gene product is a single-strand DNA binding protein that promotes annealing. The gam gene product inhibits the RecBCD nuclease preventing linear DNA (i.e. PCR product) degradation.
XXFeatures:


• Homologous Recombination-based technology to modify the host chromosome
• Use PCR or donor cell DNA to directly and precisely alter plasmids, BACS, or host genomic DNA
• Arabinose-inducible exo, bet, and gam and orf 60a genes
• Conditional temperature sensitive (ts) vector that is maintained at 30ºC, and lost at 42ºC
• Counter-selection against plasmid in streptomycin resistant (rpsL-based) host strains (pKD46-recA)
• Polylinkers for addition of new genes into vector (pKD46-recApa)
XXVector Specifications:


pKD46-recA encodes constitutively expressed Escherichia coli recA+ along with wild type rpsL. Plasmid borne rpsL converts streptomycin resistant rpsL strains such as DH10B into streptomycin sensitive strains, allowing counterselection-based plasmid elimination (Imam et al., 2000). 

DNA Vaccine Vector, pKD46-recA

pKD46-recApa encodes constitutively expressed Pseudomonas aeruginosa recA+. The RecApa protein induces hyper recombination in E. coli, in the absence of SOS induction, and presence or absence of E. coli RecA protein

DNA Vaccine Vector, pKD46-recApa

Both vectors have been validated for use in PCR- or genomic DNA-mediated recombineering applications.