Cloning of the human interleukin-23 receptor extracellular region gene and its expression in Escherichia coli.

Objective To clone the human interleukin-23 receptor(hIL-23R) extracellular domain gene and express the gene in Escherichia coli BL21(DE3). Methods The hIL-23R extracellular fragment was amplified by PCR.The recombinant plasmid pGEX-4T-1-hIL-23R(O) was constructed by cloning the extracellular fragment of hIL-23R into the prokaryotic expression vector pGEX-4T-1.After the recombinant plasmid was identified by restriction endonuclease digestion analysis and DNA sequencing,pGEX-4T-1-hIL-23R(O) was transformed into E.coli BL21(DE3) through IPTG induction to express the target protein.The expressed protein was analyzed by SDS-PAGE and Western blot.Results A 990 bp extracellular fragment of the hIL-23R gene was obtained by PCR,inserted into the prokaryotic expression vector pGEX-4T-1-hIL-23R(O) and expressed in E.coli BL21(DE3).SDS-PAGE showed the molecular weight of the fusion protein to be 64 ku,and Western blot showed that the fusion protein could be recognized by the specific rabbit anti-GST antibody.Conclusion The hIL-23R extracellular domain recombinant expression plasmid pGEX-4T-1-hIL-23R(O) was successfully constructed,and the fusion protein GST-hIL-23R was successfully expressed in E.coli BL21.This research lays the foundation for further functional study of hIL-23R.