CpG-pVAX1-ORF5真核重组表达质粒小鼠细胞免疫的研究  

Study on Cellular Immune Response Evicited Eukaryotic Expression Vector CpG-pVAX1-ORF5

作　　者：温建新[1] 徐文[1] 王金宝[2] 

机构地区：[1]青岛农业大学动物科技学院 [2]山东省农业科学院

出　　处：《青岛农业大学学报（自然科学版）》2008年第2期95-100,共6页Journal of Qingdao Agricultural University(Natural Science)

基　　金：山东省自然科学基金项目资助(编号:Z2003D01)

摘　　要：为了提高猪繁殖与呼吸综合征病毒(PRRSV)基因免疫的效果,构建含CpG基序和PRRSV ORF5的真核重组表达质粒CpG-pVAX1-ORF5。经酶切鉴定和序列测定,将重组质粒转染COS-7细胞,经间接免疫荧光试验证实CpG-pVAX1-ORF5可表达PRRSV GP5蛋白。免疫SPF小鼠,检测鼠的脾T淋巴细胞亚群数量(CD4+和CD8+)以及淋巴细胞增殖反应(MTT法)水平。试验结果表明,本试验构建的含CpG基序真核重组表达质粒CpG-pVAX1-ORF5能诱导小鼠产生针对PRRSV的细胞免疫。To improve the effect of the gene immunization against porcine reproductive and respiratory syndrome virus （PRRSV）we constructed the eukaryofic expression vector CpG -pVAX1 - ORF5 containing PRRSV ORF5 gene coding region and CpG motif. After identified by enzyme analysis and nucleotide sequencing test , the recombinant expression vector plasmid CpG- pVAX1 -ORF5 was transfected into COS-7 cells. The transient expression of PRRSV ORF5 nucleocapsid protein was detected by indirect immunofluorescence assay（IFA）. The recombinant expression plasmid was inoculated into SPF mice. The number of CD^4＋, CD^8＋ and the lymphocyte proliferation test （ MTT ） in peripheral blood of mice vaccinated were used to detect. The results showed that the recombinant plasmid CpG - pVAX1 - ORF5 could induce higher response of cellular immune response.

关 键 词：猪繁殖与呼吸综合征病毒 ORF5基因 细胞免疫应答 

分 类 号：S852.65[农业科学—基础兽医学]