这研究被设计在象蛋白质 2 一样的鼠标纤维蛋白原(mfgl2 ) 的表达式的抑制探索核糖核酸干扰技术,它被报导了涉及开发包括暴发性的病毒的肝炎的许多疾病。一个原生质标志说出 p-mfgl2shRNA,对 mfgl2 的顺序补足被构造,当是 mfgl2shRNA 序列的一种变异的形式的另一短发卡核糖核酸(shRNA ) 被用作控制时。表示 mfgl2-EGFP 熔化蛋白质的命名 pEGFP-mfgl2 也是的一个原生质标志为在 mfgl2 表示上屏蔽 p-mfgl2shRNA 的效果构造了。由 p-mfgl2shRNA 的 cotransfection;pEGFP-mfgl2 或 pcDNA3.1-mfgl2 表示构造进 CHO 房间或 HeLa 房间,由 mfgl2shRNA 的 mfgl2 表示的抑制被直接观察通过荧光灯的显微镜学分析, FACS, RT-PCR;染色的免疫组织化学。实验在两 CHO 在 48h post-transfection 在 mfgl2 表示上显示出 p-mfgl2shRNA 的重要抑制酌;有象 80.1% 一样高的禁止的效率的 Hela 细胞株。学习证明 p-mfgl2shRNA 的构造成功地防碍 mfgl2 表示离体。
This study was designed to explore the RNA interference technique in inhibition of the expression of the mouse fibrinogen like protein 2(mfgl2),which has been reported to be involved in the development a variety of diseases including fulminant viral hepatitis.A plasmid named p-mfgl2shRNA,complementary to the sequence of mfgl2 was constructed,while another short hairpin RNA(shRNA) which was a mutated form of the mfgl2shRNA sequences was used as a control.A plasmid named pEGFP-mfgl2 expressing the mfgl2-EGFP fusion protein was also constructed for the screening of the effect of p-mfgl2shRNA on mfgl2 expression.By cotransfection of p-mfgl2shRNA and pEGFP-mfgl2 or pcDNA3.1-mfgl2 expression construct into CHO cells or HeLa cells,the inhibition of mfgl2 expression by mfgl2shRNA was analyzed by direct observation through fluorescent microscopy,FACS,RT-PCR and immunohistochemistry staining.The experiments showed the significant inhibitory effect of p-mfgl2shRNA on mfgl2 expression at 48h post-transfection in both CHO and Hela cell lines with the inhibitory efficiency as high as 80.1%.The study demonstrated that the construct of p-mfgl2shRNA successfully interfered with the mfgl2 expression in vitro.