Construction and identification of prokaryotic expression vector of human cystatin C / 重庆医科大学学报

ABSTRACT

Objective:To Construct a prokaryotic expression vector of cystatin C(Cys C)to provide a basis of purify Cys C protein produced by the expression system.Methods:Total RNA was isolated from HL-60 cells,and human Cys C gene was amplified with RT-PCR.The cDNA fragment was cloned into pET32a(+)vector,which was confirmed with sequencing and electrophoresis.Results:The result of electrophoresis showed that Cys C had been cloned into pET32a(+)vector,and the result of.DNA sequence analysis also showed the cloned Cys C gene sequence was completely correspondent to GeneBank data.Conclusion:The prokaryotic expression vector of cystatin C(Cys C)was obtained,providing a basis of purification of Cys C protein produced by the expression system.