With the support of our advanced protein engineering platform established for many years, Creative BioMart has developed a variety of homologous recombination methods. Our scientists provide you with a comprehensive customized homologous recombination library construction service.

Homologous recombination introduces a vector containing a DNA sequence similar to the gene to be modified into the cell, and replaces the target gene in the chromosome through the process of recombination. Homologous recombination provides high-fidelity, template-dependent repair or tolerance to complex DNA damage, including DNA gaps, DNA double-strand breaks (DSB), and cross-links between DNA strands, and plays a key role in maintaining genome stability. In addition, homologous recombination proteins protect stalled replication forks as well as recover stalled or broken forks.

Protein engineering with homologous recombination develops chimeric proteins by exchanging fragments between two parental proteins. Compared with other mutagenesis techniques, this technique can introduce a high degree of sequence diversity while retaining the protein's ability to fold into its tertiary structure or three-dimensional shape and important structural interactions.

Fig 1. Mechanism of Homologous Recombination. (Jiang Y, et al., 2018)

Methods of Homologous Recombination

• Yeast recombination cloning: this method is very effective, only 29 nucleotides of overlapping sequence can be added to the synthetic oligonucleotide.
• Recombination engineering: it allows the insertion, deletion or modification of any sequence, independent of the position of the restriction site, and is catalyzed by the homologous recombination function encoded by the phage.
• Single-strand annealing: there is a homologous sequence at each end of the double-strand break, and the homologous sequence region is directly annealed to form a complementary double-strand, and then the double-strand break is repaired through end processing and connection.
• Synthesis-dependent annealing: this approach is a repair process that relies on DNA synthesis. Homologous recombination is usually carried out in this way during genome editing.
• DNA shuffling: it is a method of in vitro recombination of homologous genes, which can produce directed evolutionary mutant genes, and can quickly increase the size of the DNA library.

Applications of Homologous Recombination

• Specific mutations in the organism.
• Gene knockout.
• Generating new DNA sequence combinations.
• DNA repair.
• Designing a new protein.
• Enzyme Engineering.

Service Principle

Creative BioMart is based on innovative, pragmatic and honest, adhering to the tenet of "Quality is our life, providing customers with the best quality service", providing customized services to customers around the world.

We will be glad to discuss details of intended interaction studies with you and develop experimental strategies/methods tailored to your requirement. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, Monday to Friday. Please do not hesitate to contact us for more information or to discuss in detail, if you are interested in our services.

References

1. Li, X., Heyer, WD. Homologous recombination in DNA repair and DNA damage tolerance. Cell Res 18, 99–113 (2008).
2. Kim SY, Sohn JH, et al.. (2003) Efficient library construction by in vivo recombination with a telomere-originated autonomously replicating sequence of Hansenula polymorpha. Appl Environ Microbiol. 69(8): 4448-54.
3. Jiang Y, Chu WK. (2018) Potential Roles of the Retinoblastoma Protein in Regulating Genome Editing. Front Cell Dev Biol. 6:81.
4. Sharan SK, Thomason LC, et al.. (2009) Recombineering: a homologous recombination-based method of genetic engineering. Nat Protoc. 4(2): 206-23.

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