肝细胞生长因子在LO_2细胞表达的实验研究

Experimental study of hepatocyte growth factor recombinant gene expression in LO_2 cell line

目的 将肝细胞生长因子 (hepatocytegrowthfactor ,HGF)真核表达载体 ,转染培养的LO2 肝细胞 ,观察其表达。方法 将含肝细胞生长因子cDNA真核表达载体PEGFP C1 HGF ,脂质体 (Lipo fectamine)转染培养的LO2 肝细胞 ;PCR鉴定外源基因的导入 ,绿荧光蛋白检测肝细胞生长因子的表达。结果 ( 1)重组质粒PEGFP C1 HGF转染LO2 肝细胞 ,5d后 ,细胞内可检测到绿荧光蛋白。 ( 2 )PCR结果显示 :转染PEGFP C1 HGF质粒组可见 3 0 6bp特征条带与阳性对照大小一样 ,假转染组与空质粒组无特征条带。结论 人肝细胞生长因子基因真核表达载体 ,PEGFP C1 HGF ,能成功转染LO2

Objective To examine the expression of hepatocyte growth factor in the LO 2 cell transfected with constructed expression plasmid containing the HGF cDNA.Methods The recombined eukarvotic vector, inserted with hepatocyte growth factor cDNA,PEGFP C1 HGF,transfected the culture cell LO 2 hepatocyte with lipofectamine.The inserted DNA was identified by PCR.The expression of hepatocyte growth factor protein was determined by measuring green fluorescent protein.Results (1) after four days of transfection with recombined PEGFP C1 HGF,the green fluorescent protein can be observed in the L0 2 cell.(2) the result of PCR showed the group transfected with PEGFP C1 HGF was characterized with the 306bp fragment,same size as the positive control,however,the group of pesu transfection and negative group had no character fragment.Conclusion Human eukarvotic expression vector,PEGFP C1 HGF,containing green fluorescent protein gene can successfully transfect LO 2 cell and express in it.