A novel method for gene-targeting vector construction—Red/ET recombination |
| |
Authors: | Jian-Ling YANG Shu-Ping GU Chen CHEN Zhu-Gang WANG Yan XU Jian FEI |
| |
Affiliation: | aDepartment of Life and Environmental Sciences, Shanghai Normal University, Shanghai 200234, China;bShanghai Research Center for Biomodel Organism, Shanghai 201203, China |
| |
Abstract: | The study of the function of novel human genes has become an increasingly active academic field with the gene-knockout (KO) mouse model forming the basis of this study. Because of the low efficiency of recombination of targeting vector constructed using the traditional method, the KO mouse model has become the key step in the construction of a targeting vectors, both economically and efficiently. To study the function of a novel gene (Resp18), a novel DNA engineering platform, Red/ET recombination, was introduced to construct the Resp18 targeting vector. Red/ET recombineering differs from the conventional methods of vector construction (e.g., PCR, restriction enzyme digestion, and ligation), and genetic modification is accomplished by acquisition, insertion, fusion, or replacement of the target gene through small fragments-mediated homologous recombination. At present, Resp18 targeting vectors constructed using three strategies mentioned above were successfully released through two homologous recombination processes of retrieval and neo-targeting. Red/ET recombination has the advantage of producing genes with longer homology regions without mutation. |
| |
Keywords: | Red/ET recombination knockout vector |
本文献已被 ScienceDirect 等数据库收录! |
|