Abstract: Objective To construct the prokaryotic expression vector of the human regucalcin gene.Methods To get the human regucalcin cDNA from human live cancer HepG₂ cells with RT-PCR,and then inserted the fragment of regucalcin into pMD-18 simple T vector.The amplified products were cloned into the prokaryotic expression vector pET-32a(+).Results There was one obvious band at the position of relative molecular weight 45 000 after SDS-PAGE analysis,and it was equivalent to the expected value.Conclusion To construct the recombinant fusion expression vector and to express the fusion protein in E.coli successfully.

Key words: RT-PCR, regucalcin, gene expression, fusion protein