Expression of GST-human lung carcinoma deleted gene

Objective To construct the recombinant plasmid pGEX-6p-1/human lung carcinoma deleted gene 1(HLCDG1) and to investigate the expression of HLCDG1 protein in Escherichia coli.Methods The coding region of HLCDG1 gene was obtained by polymerase chain reaction(PCR).A fusion protein expression vector pGEX-6p-1/HLCDG1 was constructed by inserting fragment of the code region of HLCDG1 gene into the fusion protein expression vector pGEX-6p-1.The coding region of HLCDG1 gene was identified by endonuclease digest and DNA sequencing.The recombinant plasmid was transformed into Escherichia coli.The expression of GST-HLCDG1 fusion protein was induced by isopropy-β-D-thiogalactoside(IPTG) and confirmed by SDS-PAGE and western blot.Results The coding region of HLCDG1 gene was correctly inserted into the pGEX-6p-1 vector and the pGEX-6p-1/HLCDG1 was constructed.After induction with IPTG,a new protein band of 46 kDa was detected on SDS-PAGE in the cells transformed with recombinant plasmid and identified by western blot. The GST-HLCDG1 fusion protein was mostly existed in the precipitation of broken bacteria.Conclusion The HLCDG1 protein can be expressed in prokaryotic expression system,which lays foundation for the further structural and functional research of the protein.