Construction And Expression Of The Prokaryotic Expression Vector Of Recombinant Human Glucocorticoid Receptor

Objective: To construct hGRa recombinant prokaryotic expression vector and induce it to express. Thiswork might serve the creating of hGRa ELISA detection kit and the further clinical study of correlationbetween glucocorticoid receptor and Yin-Yang deficiency syndrome.Methods: Two kinds of gene fragments of hGRa were amplified by PCR from the recombinant cloningplasmid pRShGRa, which carrying the hGRa cDNA sequence. After being respectively cloned into thepGEM-T vector first, the two kinds of gene fragments were subcloned into the pGEX-4T-3 expressionvector. The two kinds of recombinant expression vectors were transformed into the Escherichia coliDH5a and BL21(DE3) respectively after being analyzed by restriction endonuclease digestion andsequencing. The engineering E.coli were induced to express the fusion proteins under some inductionconditions and the induced products were observed with SDS-PAGE and Western blotting.Results: Restriction endonuclease digestion and sequencing analysis showed that the two kinds of genefragments of hGRa were recombined into pGEX-4T-3 vector successfully. The SDS-PAGE and Westernblotting analysis of the induced products showed that the two kinds of hGRa target proteins wereexpressed specifically.Conclusion: The two kinds of recombinant expression vectors were constructed and induced to expresssuccessfully, which might contribute to the following study greatly.