Recombineering Based Microbe Genome Editing

Creative Biogene is the leading company offering the best recombineering based microbe genome editing services for a large wide range of microorganism species. With the assistance of our excellent scientists and technicians, we will complete your microbe genome editing project in a shot turnaround time with most competitive price.

Creative Biogene offers microorganism genome mutation, insertion, gene knock out and metabolic pathways engineering service for a broad range of microbes by our mature recombineering platform. Our microbe targets are including but not limited to Escherichia coli, Enterobacter, Salmonella, Pseudomonas and other Gram-negative bacteria.

Recombinase-mediated homologous recombination, also known as recombineering, has revolutionized bacterial genetics, synthetic biology, and metabolic engineering. Relying on either RecET from the Rac prophage or the bacteriophage lambda Red proteins, Exo, Beta, and Gam, recombineering allows simple and efficient construction of gene knockout mutants via homologous recombination of a double stranded DNA (dsDNA) polymerase chain reaction (PCR) product with bacterial chromosomes.

To achieving the replacement of the target gene with exogenous DNA, the RecBCD system, which is widely existent in bacteria, is used in the homologous recombination. The homologous fusion fragment of the target gene was cloned into the suicide vector. Then the recombinant vector was injected into the target bacterium by conjugation. Because of the suicide vector, most bacteria can’t replicate themselves. Only the mutant strain which target fragment was integrated with genome could survive under the pressure of antibiotic selection. In the second round of reverse selection pressure, only the bacterial that have second homologous recombination and loss of suicide plasmid in genome can survive. In the second time of homologous recombination, bacteria progeny produce mutant strain and wild-type strain. By PCR screening and sequencing, we can obtain the deletion mutant of the target gene.