CAJ | 학술논문

将人肝细胞生长因子（human hepatocyte growth factor , hHGF）全长cDNA重组入pEE14真核稳定表达质粒，用lipofectin脂质体将pEE14/rhHGF转染入CHO-K1细胞，蛋氨酸亚氨基代砜（methionine sulfoximine, MSX）筛选出阳性细胞克隆。利用RT-PCR检测rhHGF mRNA的表达，通过ELISA法测定rhHGF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pEE14 /rhHGF的细胞可扩增出hHGF特异的396　bp 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rhHGF的含量在8　μg/L以上，显示rhHGF在CHO细胞中以活性形式得到表达。
장인간세포생장인자（human hepatocyte growth factor , hHGF）전장cDNA중조입pEE14진핵은정표체질립，용lipofectin지질체장pEE14/rhHGF전염입CHO-K1세포，단안산아안기대풍（methionine sulfoximine, MSX）사선출양성세포극륭。이용RT-PCR검측rhHGF mRNA적표체，통과ELISA법측정rhHGF적단백표체，3H참입법검측배양상청액대대서원대배양간세포DNA합성적영향。결과표명전염pEE14 /rhHGF적세포가확증출hHGF특이적396　bp RT-PCR편단，배양상청액명현촉진대서간세포DNA적합성，ELISA법측출상청액중rhHGF적함량재8　μg/L이상，현시rhHGF재CHO세포중이활성형식득도표체。
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.