携Hsp70的PRRSV ORF5与ORF6基因重组质粒构建及在Cos-7细胞中的表达  

Construction of eukaryotic recombinant expression vector containing Hsp70 and PRRSV ORF5,ORF6 genes and its responses in Cos-7 cell line

作　　者：左兰[1] 颜其贵[1,2] 刘彬[3] 李双江[3] 

机构地区：[1]四川农业大学动物生物技术中心,四川雅安625014 [2]四川农业大学动物疫病与人类健康四川省重点实验室,四川雅安625014 [3]四川农业大学动物医学院,四川雅安625014

出　　处：《浙江农业学报》2011年第3期495-499,共5页Acta Agriculturae Zhejiangensis

基　　金：教育部长江学者和创新团队发展计划(IRT0848);四川省教育厅重点项目(07ZA053);东莞市科技局项目(2007108101100)

摘　　要：为构建高效猪繁殖与呼吸综合征病毒(PRRS)DNA疫苗,利用Hsp70作为免疫佐剂增强免疫功能的特点,构建了含PRRSVORF5,ORF6基因的融合表达载体pCI-ORF5-ORF6A,以及携带HSP70基因的融合表达载体pCI-ORF5-ORF6B-Hsp70,转染Cos-7细胞。IFA检测结果显示,两个融合表达载体均能在Cos-7细胞中表达;Western blot检测证实转染质粒pCI-ORF5-ORF6A和pCI-ORF5-ORF6B-Hsp70在42.7 Ku,112.2 Ku分别表达出特异性条带。结果显示所构建的重组质粒能在Cos-7细胞内进行良好的表达,且表达的蛋白具有免疫原性。In order to construct efficient porcine reproductive and respiratory syndrome（PRRS） DNA vaccines and carrying HSP70 gene which can be used as an adjuvant enhancing immune function,we constructed the fusion expression vector PCI-ORF5-ORF6A and PCI-ORF5-ORF6B-HSP70 which comprehended PRRSV ORF5 and ORF6 genes.The two kinds of fusion expression vectors were transfected Cos-7 cells.IFA results showed that the two fusion expression vector in Cos-7 cells could express and western blot test confirmed that the specific bands about 42.7 Ku and 112.2 Ku could be found from transfection of plasmid pCI-ORF5-ORF6A and pCI-ORF5-ORF6B-Hsp70.It showed that the recombinant plasmid in Cos-7 cells were efficiently transcribed and expressed,and the expression of the protein was immunogenic.

关 键 词：PRRSV HSP70 COS-7细胞 

分 类 号：Q78[生物学—分子生物学]