Abstract:Objective:To construct a recombinant vector containing Tip-α of H.polyri, and transfect it in E.coli to purify recombinant protein. Methods:By PCR technique, Tip-α cDNA was amplified, and then cloned into the pET28a vector. The recombinant plasmid was transformed into E.coli. The correct clone was identified by sequence analysis. The expression product was purified by nick-nitrilotriacetic acid(Ni-NTA) metal-affinity chromatography and analyzed by SDS-PAGE and Western blot methods. Results: Enzyme digestion analysis and sequencing assay showed that Tip-α gene had been successfully inserted into the vector. SDS-PAGE showed a 23 ku protein identified by Western blot analysis. Conclusion:A recombinant plasmid containing Tip-α has been constructed and expressed successfully.