【摘要】 目的:构建人胱抑素C(Cys C)的原核表达载体,为进一步表达纯化Cys C重组蛋白奠定基础。方法:从HL-60细胞中提取总RNA,RT-PCR扩增Cys C基因,再将目的基因克隆至PET32a(+)表达载体中,构建人Cys C原核表达质粒PET32a(+) /Cys c;再将此重组子进行双酶切电泳鉴定和测序鉴定。结果:酶切电泳鉴定结果显示Cys C基因克隆入载体中,测序结果证实克隆的基因序列与GeneBank中的Cys C序列相符。结论:获得了人Cys C原核表达质粒PET32a(+)/Cys C,为进一步表达和纯化Cys C重组蛋白奠定了工作基础。更多还原

【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.更多还原