针对HO-2基因的四种RNA干扰序列构建和效应观察  

Construction and identification of four RNA interference vectors targeting on HO-2 gene

作　　者：王珍珍[1] 张腾业 陈仁金[2] 袁红花[1] 胡安康[2] 吴连连[2] 朱裕华[2] 朱孝荣[2] 

机构地区：[1]徐州医学院神经生物研究中心 [2]徐州医学院实验动物中心,徐州221004

出　　处：《神经解剖学杂志》2015年第2期161-164,共4页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(31172171);江苏省青年基金(BK2012138);徐州医学院院长基金(2012KJZ20)

摘　　要：目的:构建和筛选出血红素氧合酶-2(HO-2)基因的RNA干扰载体,并观察其在小鼠脑血管内皮细胞中的表达情况。方法:根据小鼠HO-2基因的c DNA序列设计了4个HO-2基因干扰序列,克隆到干扰载体p GPU6-GFP-Neo上,利用电穿孔法将干扰载体对小鼠脑血管内皮细胞进行转染。Real-time PCR和Western Blot检测小鼠脑血管内皮细胞中HO-2的表达。结果:干扰载体显著抑制小鼠脑血管内皮细胞中HO-2的表达,其中干扰载体p GPU6-GFP-Neo-HO-2-mus-768对HO-2 mRNA的表达抑制达显著水平(P<0.01),蛋白的表达抑制达显著水平(P<0.05)。结论:成功构建并筛选了HO-2表达干扰载体,为下一步的HO-2基因的功能鉴定奠定了基础。Objective： To construct heme oxygenase-2 interference vectors,and observe their expression in cerebrovascular endothelial cells of mice. Methods： Four interference sequences were designed according to the the c DNA sequence of HO-2 gene,which were cloned into interference vector p GPU6-GFP-Neo. Then cerebrovascular endothelial cells were transfected with the interference vector using electroporation,the Real-time PCR and Western Blot was used to detect HO-2 mRNA and protein levels in cerebrovascular endothelial cells. Results： The interference vectors highly suppressed the HO-2 gene expression of cerebrovascular endothelial cells,and the interference vector p GPU6-GFP-Neo-HO-2-mus-768 extremely significant suppressed the mRNA expression of HO-2（ P 〈 0. 01）,and the protein expression inhibition also reached significant level（ P 〈 0. 05）. Conclusion： The interference vector p GPU6-GFP-Neo-HO-2 was successfully constructed and selected,which may be benefit for further study the mechanisms and function of HO-2 gene.

关 键 词：HO-2基因 RNA干扰载体 基因表达 大鼠 

分 类 号：R741[医药卫生—神经病学与精神病学]