中文摘要：

目的 构建人肝细胞核因子-1α（hepatocyte nuclear factor1α，HNF1α）真核表达载体，在体外表达并检测。方法 提取HepG2总RNA，用RT-PCR方法制备HNF1α cDNA，进行T-A克隆。HindⅢ/EcoRⅠ双酶切测序正确的重组质粒，回收HNF1α cDNA片段，将其亚克隆于pcDNA3.1（＋）载体的多克隆位点内。将构建好的真核表达质粒用脂质体法转染HepG2细胞，RT-PCR和免疫印迹法检测HNF1α的表达。结果 正确构建了pcDNA3.1（＋）-HNF1α重组载体，并且在转染该质粒的HepG2细胞中检测出了HNF1α的高表达。结论 成功地构建了人HNF1α真核表达载体，其可以在体外表达。

英文摘要：

Objective To construct the eukaryotic expression vector of human HNF1α gene and to observe its expression in vitro. Methods Total RNA from HepG2 cell was used to acquire human HNF1α cDNA by RT-PCR. Then HNF1α cDNA was cloned into T-A vector. The recombined vector confirmed by sequencing was digested by HindⅢ/EcoR Ⅰ to obtain HNF1α cDNA fragment, then the fragment was subcloned into multiple sites of pcDNA3.1 （ ＋ ） vector. The identified recombinant plasmid was transfected into HepG2 cell with liposome. The expressions of HNF1α in the cell were determined with RT-PCR and Western blot. Results The recombinant pcDNA3.1 （ ＋ ）-HNF1α plasmid was constructed successfully and the overexpression of HNF1α gene could be detected in the transfected HepG2 cell. Conclusion The recombinant expression pcDNA3.1 （ ＋ ） -HNF1α vector is successfully constructed and can be expressed in vitro.