Expression in E.coli of protein encoded by human esophageal cancer related gene-1

Objective To express esophageal cancer related gene 1 (ECRG 1) in E.coli.Methods The human ECRG 1 was cloned by RT PCR. Expression plasmid of the ECRG 1/GST π fusion gene was constructed and introduced into E. Coli. The fusion protein was induced to express by ITPG. The recombinant protein was identified by Western blot. BALB/C mice were immunized with the protein purified by SDS PAGE for the preparation of polyclonal antibody.Results DNA sequencing confirmed that the sequence of ECRG 1 was identical to that of the previously obtained by 5′ RACE technique. Expression of the protein in E.coli was verified by Western blot. The polyclonal antibody obtained from immunized BALB/C mice had a title of 1∶3 200.Conclusion The antibody available is useful for the further study of structure and function of the esophageal cancer related protein.