Expression of Recombinant Human Thyroid Stimulating Hormone Subunit α in Escherichia Coli
10.3969/j.issn.0253-9896.2010.03.003
- VernacularTitle:重组人促甲状腺激素α亚基在大肠杆菌中的表达
- Author:
Weili XIONG
;
Bei SUN
;
Xiaona CAO
;
Jingcai MA
;
Jingyu ZHANG
;
Gang GUO
- Publication Type:Journal Article
- Keywords:
glycoprotein hormones,alpha subunit;
cloning,molecular;
recombination,genetic;
gene expression;
escherichia coli
- From:
Tianjin Medical Journal
2010;38(3):167-169
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct pGEX-3X/hTSHa Escherichia coli(E.coli)expression system and prepare purified recombinant GST-recombinant human thyroid stimulating hormone(rhTSH)α protein.Methods:The complete coding sequence of hTSHα was obtained by RT-PCR with total RNA extracted from fresh chorial tissue as the template,and thereafter cloned into expression vector pGEX-3X by EcoRl and BamHI digestion.The recombinant plasmid was transformed into E.coli Mach1-T1 and then induced expression by WrG.The GST-rhTSHα fusion protein was identified by SDS-PAGE and its antigenicity was verified by a modified competitive ELISA.Results:A specific protein band of 36 ku,in accordance with predicted molecular weight,could be visualized in SDS-PAGE.As the result of ELISA,the recombinant GST-hTSHα protein can inhibit the intact TSH molecular binding with anti-TSHα antibody in a dose dependent manner.Conclusion:The cDNA of hTSHα was cloned and the recombinant expression vector pGEX-3X/hTSHα was constructed successfully.The recombinant GST-rhTSHα protein could be highly expressed in E.coli Machl-T1 and was approved of possessing antigenicity.
