中文摘要：

为了建立稳定表达HSP72蛋白的奶牛乳腺上皮细胞系,本试验应用PCR法扩增HSP72基因,插入真核表达质粒EGFP-N2中构建EGFP-N2-HSP72重组质粒。以脂质体介导的方法将其转染于奶牛乳腺上皮细胞,经G418筛选后获得稳定表达HSP72蛋白的奶牛乳腺上皮细胞,并且转染的HSP72基因序列正确。阴性对照组EGFP-N2的奶牛乳腺上皮细胞在荧光显微镜下呈绿色荧光,且重组质粒组极显著（P〈0.01）表达HSP72蛋白。表明成功构建了EGFP-N2-HSP72真核表达载体,并建立了高表达HSP72蛋白的奶牛乳腺上皮细胞系。

英文摘要：

In order to establish a stable bovine Mammary Epithelial Cell（b MEC）line expressing HSP72 protein,HSP72 genewas amplified by PCR and inserted into the eukaryotic expression plasmid EGFP-N2 to construct the recombinant plasmid EGFP-N2-HSP72.Liposome-mediated method was used to transfect it into b MEC.After selecting by G418,the stable b MEC line expres-sion HSP72 protein was obtained.Negative control plasmid EGFP-N2 of b MEC showed green fluorescence under a fluorescence mi-croscope,and the recombinant plasmid had a very significant high expression of HSP72 protein.Here,we successfully constructedan EGFP-N2-Hsp72 eukaryotic expression vector,and established a b MEC line with high expression of HSP72 protein.