摘要
目的 构建肝细胞生长因子 (HGF)真核表达载体 ,为基因修饰肝细胞移植的细胞来源及建立 HGF的工程细胞株提供实验依据。方法 利用 DNA体外重组技术 ,将 HGF c DNA重组入真核表达载体 PEGFP- C1 ,限制性内切酶及测序鉴定聚合酶链式反应 (PCR)鉴定外源基因的导入。结果 重组真核表达载体 PEGFP- C1 -HGF经限制性内切酶分析 ,与理论值相符 ,测序结果与 Gene Bank对比 ,序列正确 ,插入正确。结论 成功构建带有绿荧光蛋白的 HGF基因真核表达载体 PEGFP- C1 - HGF。
Objective To construct eukarvotic expression plasmid containing the hepatocyte growth factor(HGF) cDNA.Thereby,it provides valuable experiment basis for the hepatocellar resource of modified hepatocyte transplantation and establishment of engineer cell line expressing HGF.Methods HGF cDNA was inserted in the eukarvotic expression vector PEGFP-C1 by DNA recombined technique in vitro,and the recombined vector,PEGFP-C 1-HGF,was identified with enzyme digestion and sequence.Results The segment of recombined expression vector PEGFP-C 1-HGF digested with enzyme corresponds to the expection.The sequence and the position inserted are accurate according to the Gene Bank.Conclusion Human eukarvotic expression vector,PEGFP-C 1-HGF,containing green fluorescent protein gene is reconstructed successfully.
出处
《山东医药》
CAS
北大核心
2003年第30期4-5,共2页
Shandong Medical Journal
