荧光蛋白融合人核糖核酸酶抑制因子基因在人肝癌细胞中的表达

摘要

Objective To investigate the expression of a retroviral vector of recombinant green fluorescent protein fused to human ribonuclease inhibitor (RI) (pLNCX-EGFP-C1-hri) in the hepatocellular carcinoma HepG2.Methods The re-combinant vector pLNCX-EGFP-C1-hri was transfected into HepG2 cells.After 2 weeks of selection of 600 mg/L G418, the expression of pLNCX-EGFP-C1-hri was detected by RT-PCR and Western blotting .Results In the HepG2 cells, the pLNCX-EGFP-C1-hri was highly expressed in the levels of gene transcription and protein (all P<0.01).Conclusion The retroviral vector of green fluorescent protein fused to human ribonuclease inhibitor is successfully constructed .%目的：探讨重组绿色荧光蛋白融合人核糖核酸酶抑制因子（ RI）的逆转录病毒载体pLNCX-EGFP-C1-hri在肝癌细胞中的表达。方法用脂质体法将所构建的重组载体pLNCX-EGFP-C1-hri转染到人肝癌HepG2细胞中，用600 mg/L G418筛选2周后采用RT-PCR和Westen blot检测pLNCX-EGFP-C1-hri表达。结果在HepG2细胞中，pLNCX-EGFP-C1-hri在基因转录水平和蛋白质水平均高效表达（P均＜0．01）。结论本研究成功构建重组绿色荧光蛋白融合人RI的逆转录病毒载体。