Cloning of human sperm protein(SP17) and expression in escherichia coli DH5?
- VernacularTitle:人精子蛋白SP17基因的克隆及其在大肠杆菌中的表达
- Author:
Minzhen WANG
;
Zhicheng LIU
;
Zhan GAO
;
Wenbo ZHENG
;
Jianhon LUO
- Publication Type:Journal Article
- Keywords:
Human;
Sperm;
Proteins;
Genes
- From:
Chinese Journal of Pathophysiology
2000;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To obtain GST fusion protein of hSP17 gene and construct the recombinant plasmid for expression in E. coli. METHODS: Total fragment of hSP17 cDNA gene were amplified by RT-PCR, then subcoloned into pGEX-3b to generate recombinant hSP17/pGEX. Right orientation of insert are identified by restricted enzyme digestion. Transform the correct recombinant plasmid into the E. coli DH5a. The expression of fusion proteins hSP17-GST were induced by adding isopropylthiogalactoside (IPTG). RESULTS and CONCLUSION: The recombinant plasmid hSP17/pGEX-3b could express effectively in E.coli and a high level of fusion protein hsp17-GST with the predicted molecular weight was detected.
