关键词: 人肝刺激因子, 原核表达载体, 基因表达

Abstract: Objective To establish the prokaryotic expression system for expression of human hepatic stimulator substance.Methods Fulllength of hHSS cDNA was digested by PstⅠand NdeⅠand the fragment of open reading frame(ORF) was inserted into pT7-7 vector.The construct of prokaryotic expression vector pT7-7-hHSS identified by endonuclease digestion and DNA sequencing.Recombinant hHSS was purified by microcon centrifugal filter.The hHSS was identified by Western blot and Peptide Mass Fingerprint.Results There was one obvious band at the position of relative molecular weight(150000),which was positively reacted with anti-HSS antibody in Western blot.The sequencing of such(150000) protein indicated that the amino acids was equivalent to the deduced data.Conclusion The recombinant fusion express vector pT7-7-hHSS is constructed,and hHSS gene is expressed in E. coli successfully.

Key words: human hepatic stimulator substance, prokaryotic expression vector, gene expression