【摘要】 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。更多还原

【Abstract】 Objective To construct the eukaryotic expression recombining vector on human hepatocyte DNA synthetic stimulated factor (hHDSSF) with gene cloning. Methods The pGEM-hHDSSF, a mid-recombining vector, was constructed by T-A cloning. After restriction endonuclease NotⅠ 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 KpnⅠdigestion 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.更多还原