Expression and purification of rice codon optimized His-cry1Ca fusion protein in E.coli and preparation of polyclonal antibody against cry1Ca

Rice codon optimized cry1Ca gene was cloned into expression vector pET-28b(+) by means of restricted enzymatic digestion.His-cry1Ca fusion protein was highly expressed by SDS-PAGE analysis accumulating more than 24% of the total bacterial proteins.The main expression production was deposited as inclusion body.His-cry1Ca was then purified with Ni-NTA after treatment of solvents.The target protein was used as the antigen to immunize rabbits.ELISA assay showed that the titer of the prepared polyclonal antibody was 1:50000,and had high specificities.