四环素诱导性人肝细胞生长因子真核表达载体的构建与鉴定

目的 构建四环素诱导性人肝细胞生长因子（HGF）真核表达载体。方法 运用基因重组技术，将人HGF cDNA全长序列，定向克隆于四环素诱导性真核表达载体pBI-L多克隆位点MluⅠ和SalⅠ之间，限制性内切酶酶切和测序鉴定。结果 重组pBI-L-HGF真核表达载体经限制性内切酶酶切，与理论值相符；测序结果与GenBank比对，序列正确。结论 本实验成功构建人HGF四环素诱导性真核表达载体pBI-L-HGF，为今后临床开展HGF基因治疗提供了一种安全可调控的基因治疗策略。

Construction and identification of tetracycline-inducible human hepatocyte growth factor eukaryotic expression vector

OBJECTIVE: To construct a tetracycline-inducible eukaryotic expression vector containing human hepatocyte growth factor (HGF) cDNA. METHODS: Human HGF cDNA fragment was obtained by PCR from pUC-SRalpha/HGF plasmid and inserted into the restriction site between Mlu I and Sal I of the tetracycline-inducible eukaryotic expression vector pBI-L. pBI-L-HGF was constructed by DNA recombination in vitro, and was identified by restriction endonucleases digestion and sequencing. RESULTS: The fragment of pBI-L-HGF digested with restriction endonucleases well corresponded to expectation, and the sequence of inserted HGF cDNA was correct according to the GenBank. CONCLUSION: The tetracycline-inducible eukaryotic expression vector of human HGF pBI-L-HGF has been constructed successfully, which allows further study of HGF gene therapy with much safety and easy manipulation.