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 pi 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.