摘要
将人肝细胞生长因子 ( human hepatocyte growth factor ,h HGF)全长 c DNA重组入 p EE1 4真核稳定表达质粒 ,用 lipofectin脂质体将 p EE1 4 /rh HGF转染入 CHO-K1细胞 ,蛋氨酸亚氨基代砜 ( methioninesulfoximine,MSX)筛选出阳性细胞克隆 .利用 RT-PCR检测 rh HGF m RNA的表达 ,通过 ELISA法测定rh HGF的蛋白表达 ,3H掺入法检测培养上清液对大鼠原代培养肝细胞 DNA合成的影响 .结果表明转染p EE1 4 /rh HGF的细胞可扩增出 h HGF特异的 396bp RT-PCR片段 ,培养上清液明显促进大鼠肝细胞 DNA的合成 ,ELISA法测出上清液中 rh HGF的含量在 8μg/L以上 ,显示 rh HGF在
The full length cDNA of human hepatocyte growth factor (hHGF) was recombinanted into pEE14 plasmid to construct eukaryotic stable expression vector. The vector was transfected into CHO K1 cells with the help of lipofectin liposome. The positive cell clone was selected under the pressure of methionine sulfoximine(MSX). RT PCR showed that positive cells expressed hHGF mRNA. The culture supernatant obtained from above system enhanced 3H thymidine incorperation into rat primarily cultured liver cells. ELISA showed that rhHGF in the culture supernatant had a concentration over 8 μg/L.
出处
《生命科学研究》
CAS
CSCD
2001年第1期60-62,共3页
Life Science Research
基金
"九五"军队重点课题基金资助项目!(96 2 0 2 5)
