Expression,purification and preliminary identification of Human HBVDNAPTP1 transactivated by hepatitis B virus DNA polymerase

Objective To construct proeukaryotic expressive vector of HBVDNAPTP1 gene,induce the expression of fusion protein in Escherichia coli. Method The DNA fragment of HBVDNAPTP1 was amplified by reverse transcription polymerase chain reaction(RT-PCR).After sequencing,the correct DNA fragment was inserted into inducible proeukaryotic expressive vector pET-32a(+).The competent BL21(DE3) E.coli was transformed,and then cultured and induced.The fusion protein of HBVDNAPTP1 was purified by the His-Binding affinity chromatography. Result The fusion protein of HBVDNAPTP1 about 31 kD was expressed optimistically by 0.5 mmol/L IPTG at 30 ℃ for 5 h and confirmed by Western Blotting.At last,2.24 mg of purified fusion protein was obtained in each litre of bacterium culture. Conclusion The purified fusion protein of HBVDNAPTP1 was obtained,which can provide a material foundation for the further study on the biologic function of HBVDNAPTP1.