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E. coli strain GB05-dir
Direct cloning-proficient E. coli strain for linear plus linear homologous recombination
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Features
- Direct cloning of DNA fragments into a linear plasmid backbone
- E. coli strain harboring an arabinose inducible ETγA operon (full length recE, recT, redγ, and recA) at the ybcC locus
Applications
- Cloning of PCR products
- Cloning of linear DNA fragments from digested genomic DNA (Fu et al. 2012)
Description
Full-length Rac prophage protein RecE and its partner RecT mediate highly efficient linear-to-linear homologous recombination mechanistically distinct from conventional recombineering mediated by Redαβ from lambda phage. E. coli strain GB05-dir enables direct cloning of DNA fragments without the necessity to maintain an expression plasmid in the cells since the necessary genes are located on the chromosome.
Contents
- Glycerol stock of direct cloning-proficient E. coli strain GB05-dir
- Linear control plasmid backbone (ColE1 origin of replication and an ampicillin resistance marker gene)
- Linear control insert (PCR product)
- Protocol
