Recombineering Based Microbe Genome Editing Service

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.

Fig.1 Overview of bacteriophage lambda Red recombination system used for recombineering (Shyam, et al. 2009)

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.

Fig.2 The Schematic of Homologous Recombination mediated by RecBCD

Creative Biogene provides a range of microorganism genome modification services, which is including but not limiting to follows:

Genome mutation: especially gene site-directed mutation.

Gene Knockout: including non-housekeeping and nonlethal gene deletion, multiple gene deletion, and chromosomal deletion and so on. Creative Biogene recombination platform allows deletions of one to several genes, entire chromosomal regions or point mutations in a scarless way (no residual nucleotide).

Gene Knock In: including different sequences or tags (like GFP, mCherry, ect.) upstream or downstream of targeted genes.

Metabolic pathways engineering: including insertion of novel metabolic pathways, modification of endogenous pathways to produce compounds that microorganisms do not express naturally. By designing and inserting completely new metabolic pathways, a microorganism will be able to acquire new functions. Creative Biogene helps our partners in selecting the strain, designing the pathways, optimizing it, validating it and delivers a ready-to-use strain.

With years of professional and abundant experience in this field, Creative Biogene can provide you an advanced genome editing service with our mature recombineering platform, quick turnaround time, and uncompromising quality of products. Modifications from one to many can also be completed by Creative Biogene on demand of your provided strains. Creative Biogene has confidence in achieving a full range of R&D studies by close collaboration with our customers to make sure all the options in obtaining the desired strain.

If you have any special requirements in recombineering based microbe genome editing service, please feel free to contact us. We are looking forward to working together with your attractive projects.

Reference:

1. Shyam, K. S., Lynn, C. T., Sergey, G., Donald, L. C. (2009) "Recombineering: A Homologous Recombination-Based Method of Genetic Engineering". Nat Protoc., 4(2), 206–223.

* For research use only. Not intended for any clinical use.

Quick Inquiry