摘要
本研究从牛外周血单个核细胞提取细胞总RNA,RT-PCR获得编码牛PU.1蛋白的cDNA;以pIRES2DsRed-Ex-press2为骨架构建牛PU.1基因表达载体pIRES2DsRed-Express2-PU.1,并利用脂质体介导重组质粒分别转染H293T细胞和牛胎儿成纤维细胞,通过荧光观察和Western blotting鉴定表明成功构建了牛PU.1的表达载体,为下一步研究牛血单核细胞分化及牛抗病育种奠定基础。
Total RNA was extracted from bovine,the cDNA encoding PU.1 protein was cloned using reverse transcription polymerase chain reaction(RT-PCR).The PU.1 gene was subcloned into pIRES2 DsRed-Express2 vector and then the recombinant plasmids were transferred into H293T cells and bovine fetal fibroblasts cells by the method of lipofectaming.The results showed that we had successfully constructed the expression plasmid of PU.1 gene by fluorescence observation and identification of Western blotting,which layed the foundation for the differentiation of bovine blood monocytes and breeding for disease resistance in cattle.
出处
《中国畜牧兽医》
CAS
北大核心
2012年第11期69-72,共4页
China Animal Husbandry & Veterinary Medicine
基金
优质肉牛新品种(系)选育与关键技术研究及示范(2011BAD28B04)
基于高密度SNP建立中国肉牛群体全基因组选择优化方案(2010jc-2)