稳定表达猪圆环病毒2型衣壳蛋白P815细胞系的建立  

Establishment of P815 cell line stably expressing porcine circovirus type 2 capsid protein

作　　者：宋勤叶[1] 杨汉春[1] 郭鑫[1] 盖新娜[1] 

机构地区：[1]中国农业大学农业部预防兽医学重点开放实验室

出　　处：《中国兽医学报》2008年第6期646-648,662,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30400321)

摘　　要：应用脂质体介导的基因转染方法,将表达猪圆环病毒2型(porcine circovirus,PCV2)衣壳蛋白的重组质粒pcDNA3.1-orf2转染P815细胞,筛选G418抗性单克隆细胞,并经PCR、RT-PCR和间接免疫荧光鉴定,随后对目的蛋白表达稳定性进行了检测。结果表明PCV2orf2基因已经整合进P815细胞基因组DNA中,ORF2编码的衣壳蛋白在获得的阳性P815细胞中稳定表达,建立了稳定表达PCV2衣壳蛋白的P815细胞系。该细胞系的建立为PCV2新型疫苗的研究提供了必要的实验材料,同时也可为其他病毒疫苗的研究提供有价值的参考。Recombinant plasmids pcDNA3.1-orf2 expressing PCV2 capsid protein were transfected into P815 line cells mediated by liposomes.The positive anti-G418 monoclonal cells were screened and obtained in the medium with G418,and identified by PCR,RT-PCR and indirect immunofluorenscence assay.Subsequently,the stability of the positive cells expressing aimed protein was detected.The results showed that orf2 gene integrated into the genome of P815 cells,the capsid protein coded by PCV2 ORF2 was expressed in the acquir...

关 键 词：PCV2衣壳蛋白 P815细胞系 表达 CTL杀伤活性 

分 类 号：S852.65[农业科学—基础兽医学]