金黄色葡萄球菌聚集因子A基因与牛IL-18基因真核共表达载体的构建与表达  

Construction of eukaryotic co-expression plasmid of pBUDCE/ClfA/BoIL-18 and its expression

作　　者：陈智[1] 刘少宁[1] 曹丙蕾[1] 孔娜[1] 李宏梅[1] 胡敬东[1] 赵宏坤[1,2] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]青岛农业大学食品科学与工程学院,山东青岛266109

出　　处：《中国兽医学报》2010年第6期766-769,788,共5页Chinese Journal of Veterinary Science

基　　金：教育部高校科技创新工程重大培育资金项目(CNO706039)

摘　　要：参照聚集因子A(Clumping factor A,ClfA)和牛白介素18(BoIL-18)cDNA序列分别设计引物进行扩增,将ClfA基因和BoIL-18基因分别插入到真核双表达载体pBUDCE4.1的CMV和EF-1α2个启动子的下游,构建携带2个目的基因的重组真核双表达载体pBUDCE/ClfA/BoIL-18。在Cellfectin Reagent的作用下转染293T细胞,用间接免疫荧光试验检测ClfA和BoIl-18蛋白。结果表明,重组质粒pBUDCE/ClfA/BoIL-18在293T细胞中同时表达了ClfA和BoIL-18蛋白。该重组载体的成功构建为研究ClfA和BoIL-18重组共表达产物的生物学活性及其防治奶牛金黄色葡萄球菌性乳腺炎的作用奠定基础。To detect the co-expression of clumping factor A（ClfA） gene and bovine interleukin-18 （BoIL-18） gene,the recombinant co-expressed vector of pBUDCE/ClfA/BoIL-18 was constructed.ClfA gene and BoIL-18 gene were amplified by two pairs of specific primers and cloned into pBUDCE4.1 at the downstream of promoter CMV and EF-1α.The recombinant vector was transfected into 239T cell by Cellfection reagent.Indirect immunofluorescence assay （IFA） showed that the recombinant vector co-expressed ClfA protein and BoIL-18 protein.The construction of this recombinant plasmid lays a foundation to further study the effect of BoIL-18 immunologic activity and represents a new type of vaccine against S.aureus infection.

关 键 词：金黄色葡萄球菌 乳腺炎 聚集因子A 牛白介素18 真核表达 

分 类 号：S852.61[农业科学—基础兽医学]