Aim: To express a candidate hEra binding protein A19 in Escherichia coli and to prepare anti-A19 antibody.
Methods: A19 gene was amplified by PCR from the plasmid containing A19 gene and was cloned into the expression vector pGEX-4T3 which was then transformed into E.coli. The A19 protein was expressed under IPTG induction. Antiserum was prepared by immunizing rabbits with the expressed A19 protein. The titer and specificity of polyclonal antibody were detected by Western blot.
Results: The expressed A19 accounted for about 30.2% of total bacterial protein. The titer of the antiserum was about 1:4 000. Western blot analysis indicated that the antiserum had high specificity.
Conclusion: A19 fusion protein was highly expressed. The specific anti-A19 antiserum was prepared successfully.