绵羊干扰素调节因子-1基因在COS-7细胞中转染表达的研究  

Study on Expression of Sheep Interferon Regulatory Factor-1 in Transfected COS-7 Cells

作　　者：王玳玮[1] 赖慧颖[1] 李玉林[1] 杨柳[1] 靳薇[1] 连正兴[1] 邓学梅[1] 

机构地区：[1]中国农业大学动物科技学院,畜禽育种国家工程实验室,北京100193

出　　处：《中国畜牧杂志》2012年第3期6-9,共4页Chinese Journal of Animal Science

基　　金：转基因生物新品种培育重大专项抗病转基因羊新品种培育(2008zx08008-005)

摘　　要：本研究旨在构建真核表达载体pIR ES2-EGFP-IR F1,并在COS-7细胞中过量表达。根据绵羊IRF1基因cDNA序列设计、合成引物,构建真核表达载体pIR ES2-EGFP-IR F1质粒,用电穿孔法,以表达质粒转染COS-7细胞,并用LPS刺激,用定量PCR方法检测IRF1基因的表达量是否升高。结果表明:重组质粒pIR ES2-EGFP-IR F1转染细胞后,可以观察到绿色荧光蛋白的表达,并且可以过量表达,与空白对照组相比差异极显著(P<0.01),说明通过转染的确会对细胞造成影响,排除转染试剂及未融合GFP的作用后,效果仍然明显。利用LPS刺激后,随着作用时间延长至24 h,IRF1基因的表达量较未用LPS作用的对照组有更高倍数的提高。To construct an expression plasmid pIRES2-EGFP-IRFI, and to overexpress it in COS-7 cells, sheep IRFI gene was cloned by PCR, and then the expression plasmid pIRES2-EGFP-IRFI was constructed. The constructed plasmid was transfected into COS-7 ceils by electroporation method. The expression of IRFI was detected by the Real- time PCR. In this study, expression plasmid pIRES2-EGFP-IRFI, and IRFI gene was overexpressed in the COS-7 cells at both conditions of stimulation by LPS and non-stimulation. The result can give base to further study of producing transgenic sheep or other animals.

关 键 词：IRF1 基因转染 过量表达 COS-7细胞 绵羊 

分 类 号：S826.2[农业科学—畜牧学]