New option for gene-targeting vector construction Red/ET recombination.
- Author:
Jian-Ling YANG
1
;
Shu-Ping GU
;
Chen CHEN
;
Zhu-Gang WANG
;
Yan XU
;
Jian FEI
Author Information
1. Department of Life and Evironment Sciences, Shanghai Normal University, Shanghai, China.
- Publication Type:Journal Article
- MeSH:
Bacteriophage lambda;
genetics;
Chromosomes, Artificial, Bacterial;
genetics;
Gene Targeting;
methods;
Genetic Engineering;
methods;
Genetic Vectors;
genetics;
Plasmids;
genetics;
Viral Proteins;
genetics
- From:Chinese Journal of Biotechnology
2006;22(6):919-924
- CountryChina
- Language:Chinese
-
Abstract:
The study of human new gene's function has become an increasingly active academic field basically relying on the gene knockout (KO) mouse. The construction of targeting vector economically and efficiently has turned into the key step to acquire a KO mouse because of the low efficiency of recombination with traditional constructed targeting vector. For study of the function of new gene-Resp18, we brought in a new DNA engineering platform-Red/ET recombination to construct Resp18 targeting vector. Red/ET recombineering differs from the conventional ways of vector construction (e.g., PCR, restriction enzyme digestion and ligation) and achieves genetic modification by acquisition, insertion, fusion or replacement of the target gene through small fragments mediated homologous recombination. Now Resp18 targeting vectors of three strategies were yielded successfully through two homologous recombination processes of retrieve and neo-targeting. Red/ET recombination has the advantage of getting longer homology regions without mutation, which makes it a new and reliable alternative to the construction of a targeting vector today.
