南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (10): 1443-1445.

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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

任淑婷； 于琳华； 徐长福； 高广道；   

  1. 西安交通大学医学院病理学系； 西安交通大学医学院生理学与病理生理学系 陕西西安710061； 陕西西安710061；
  • 出版日期:2006-10-20 发布日期:2006-10-20
  • 基金资助:
    西安交通大学博士论文基金项目(DFXJTU2004-14)~~

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

REN Shu-ting1, YU Lin-hua1, XU Chang-fu1, GAO Guang-dao2 Department of Pathology1, Department of Physiology and Pathophysiology2, Medical School of Xi’an Jiaotong University, Xi’an 710061, China   

  1. 西安交通大学医学院病理学系； 西安交通大学医学院生理学与病理生理学系 陕西西安710061； 陕西西安710061；
  • Online:2006-10-20 Published:2006-10-20

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摘要： 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HGFcDNA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。 

Abstract: 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-SRα/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.

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