人源性肝细胞生长因子高效真核表达载体的构建

目的：通过基因克隆构建人源性肝细胞生长因子(hHDSSF)真核高效表达重组体。方法：通过T-A克隆构建hHDSSF中间重组体pGEM-hHDSSF；酶切鉴定后构建真核表达重组体pcDNA3．1hisB-hHDSSF；末端终止法测定插入片断基因序列。结果：酶切证实hHDSSF完整插入pGEM中；酶切筛选得到正向插入的真核表达重组体pcDNA3．1hisB-hHDSSF，并经序列分析证明。结论：成功构建了真核表达重组体pcDNA3．1hisB-hHDSSF，为进一步转染真核细胞建立稳定的转染表达细胞株和高效表达hHDSSF奠定了坚实的基础。

Construction of high efficient eukaryotic expression recombinant on human hepatocyte DNA synthetic stimulated factor]

OBJECTIVE: To construct the eukaryotic expression recombining vector on human The pGEM-hepatocyte DNA synthetic stimulated factor (hHDSSF) with gene cloning. METHODS: hHDSSF, a mid-recombining vector, was constructed by T-A cloning. After restriction endonuclease Not I digestion, the target fragment was subcloned into eukaryotic vector pcDNA3. 1hisB to construct the eukaryotic expression recombinant pcDNA3. 1hisB-HDSSF. RESULTS: The forward insert recombinant pcDNA3. 1hisB-HDSSF was screened and obtained with restriction endonuclease Kpn I digestion and it was detected by DNA sequence analysis. CONCLUSION: The eukaryotic expression recombinant pcDNA3. 1hisB-hHDSSF on hHDSSF is constructed successfully, which lays a foundation for building a stable eukaryotic expression cell strain and expressing hHDSSF.