中国人兽共患病学报2012，Vol.28Issue(9)：947-950,4.DOI:10.3969/cjz.j.issn.1002-2694.2012.09.018
弓形虫ROP5基因真核表达载体的构建及其免疫保护性的研究
Construction of eukaryotic expression vector of Toxoplasma gondii ROP5 gene and its immunoprotective effect
摘要
Abstract
The purpose of this study was to construct eukaryotic expression pcDNA-ROP5 and evaluated the immune responses induced by recombinant plasmid pcDNA-ROP5 of T. gondii in Kunming mice. The gene sequence encoding ROP5 of T. gondii was amplified by PCR and inserted into the eukaryotic expression vector pcDNA-3. 1, and the recombinant plasmid pcDNA-ROP5 was constructed. Then the recombinant plasmid pcDNA-ROP5 was transfected into Hela cells to test its expression and the recombinant protein characterized by Western blotting. Forty-five mice were divided into 3 groups randomly; pcD-NA-ROP5 as experimental group, and pcDNA3. 1 and NS as control. Each mouse was injected intramuscularly by 100 μg recombinant plasmid for 3 times every two weeks. Mice were bled on day 0, 13 , 27 and 41 for 5 mice of each group to detect the level of IgG. And the rest mice were challenged with 1 000 tachyzoites of the virulent T. gondii RH strain at the 14th day after the last immunization to observe the survival time. In this study, the recombinant plasmid pcDNA-ROP5 was successfully constructed by PCR and restriction analysis, and the ROP5 gene of T. gondii was successfully expressed in Hela cells (Mr. 61 kDa). The pcDNA-ROP5 vaccine could induce humoral immunity in Kunming mice. The level of IgG antibodies in pcDNA-ROP5 vaccine injected mice serum was higher than those in mice with other groups. After infected with Toxoplasma tachyzoi-tes,the survival time of pcDNA-ROP5 immunized mice in experimental group was prolonged comparing with control groups. The ROP5 of T. gondii could trigger immuno-protection against T. gondii challenge in Kunming mice.关键词
弓形虫/ROP5/真核表达质粒/免疫保护Key words
Toxoplasma gondii /rhoptry protein5(ROP5) / immuno-protection分类
医药卫生引用本文复制引用
赵焕阁,王华,黄用豪,周松林,郭峻莉,黄凤迎,林映莹,谭光宏..弓形虫ROP5基因真核表达载体的构建及其免疫保护性的研究[J].中国人兽共患病学报,2012,28(9):947-950,4.基金项目
海南省自然科学基金(No.809018)资助 （No.809018）
