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Expression of Recombinant Human Thyroid Stimulating Hormone Subunit α in Escherichia Coli / 天津医药
Tianjin Medical Journal ; (12): 167-169, 2010.
Article in Chinese | WPRIM | ID: wpr-472695
ABSTRACT

Objective:

To construct pGEX-3X/hTSHa Escherichia coli(E.coli)expression system and prepare purified recombinant GST-recombinant human thyroid stimulating hormone(rhTSHprotein.

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.

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Tianjin Medical Journal Year: 2010 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Tianjin Medical Journal Year: 2010 Type: Article