猪Mx1基因真核表达载体的构建、鉴定及其表达

Construction,Identification and Expression of Eukaryotic Expression Vector of Pig Mx1 Gene

为获得转Mx1基因的阳性陆川猪成纤维细胞,本研究以干扰素诱导猪成纤维细胞Mx1基因表达,提取细胞总RNA,RT-PCR获得编码猪Mx1蛋白的cDNA;以pMSCV-IRES-GFP为骨架构建猪Mx1基因表达载体pMSCV-IRES-GFP-Mx1,并利用脂质体2000介导重组质粒转染陆川猪胎儿成纤维细胞,通过荧光观察和PCR检测分析结果表明Mx1蛋白基因整合进入陆川猪胎儿成纤维细胞。

The purpose of this study was to obtain pig fibroblast cells that transferred Mx1 gene.Total RNA was extracted from pig fibroblast cells that induced with IFN and the cDNA encoding pig Mx1 protein was cloned using reverse transcription polymerase chain reaction（RT-PCR）.The Mx1 gene was subcloned into pMSCV-IRES-GFP vector and then the recombinant plasmids were transferred into Luchuan pig fibroblast cells by the method of Lipofectaming 2000.The results showed that Mx1 gene had been integrated into the genome of Luchuan pig fibroblast cells based on the analysis of fluoroscopy and PCR identification.