弓形虫主要表面抗原p30单价及复合基因疫苗的构建  

Construction of Monovalent and Compound Nucleic Acid Vaccines Against Toxoplasma gondii with Gene Encoding p30

作　　者：杨婷婷[1] 何深一[1] 蒋华[1] 古钦民[1] 丛华[1] 周怀瑜[1] 张加勤[1] 李瑛[1] 赵群力[1] 

机构地区：[1]山东大学医学院寄生虫学教研室,济南250012

出　　处：《中国寄生虫学与寄生虫病杂志》2005年第1期14-17,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金 (No 30 371 2 57) ;教育部留学回国人员科研启动基金 (教外司留 [2 0 0 3] 4 0 6号 )~~

摘　　要：目的　构建弓形虫单价基因疫苗 pcDNA3 1 p3 0及复合基因疫苗 pcDNA3 1 p3 0 ROP2 ,并比较两种疫苗对小鼠的免疫保护性。　方法　用聚合酶链反应 (PCR)从弓形虫RH株基因组DNA中分别扩增编码弓形虫主要表面抗原 p3 0和弓形虫棒状体蛋白 2 (ROP2 )的基因片段 ,经T A克隆 ,将p3 0单价基因及 p3 0 ROP2复合基因片段分别插入真核细胞表达载体pcDNA3 1,构建重组真核表达质粒 pcDNA3 1 p3 0及pcDNA3 1 p3 0 ROP2。分别免疫BALB/c小鼠 ,设磷酸缓冲盐溶液 (PBS)组、pcDNA3 1空质粒组为对照 ;酶联免疫吸附测定 (ELISA)检测血清特异性IgG抗体 ;弓形虫速殖子腹腔攻击感染观察小鼠生存时间。 　结果　获得 pcDNA3 1 p3 0、pcDNA3 1 p3 0 ROP2重组表达质粒 ,用 pcDNA3 1 p3 0 ROP2免疫的小鼠 ,其IgG抗体吸光度 (A490 =2 0 5 1± 0 3 3 7)高于用 pcDNA3 1 p3 0的吸光度 (A490 =1 892± 0 3 69) (P <0 0 5 )。攻击感染弓形虫后小鼠生存时间 ,用 pcDNA3 1 p3 0 ROP2免疫的小鼠 ,较用 pcDNA3 1 p3 0的明显延长 (P <0 0 1)。 　结论　弓形虫不同生活阶段的抗原复合基因疫苗较单价基因疫苗具有更好的免疫保护性。Objective To construct a monovalent gene vaccine pcDNA3 ^1-p30 and a compound gene vaccine pcDNA3 ^1-p30-ROP2 and assess the protective effect of the two vaccines against Toxoplasma gondii. Methods The sequences encoding p30 and ROP2 were amplified from the genomic DNA of T.gondii RH strain by polymerase chain reaction (PCR) and inserted into eukaryotic vector pcDNA3 ^1 to construct pcDNA3 ^1-p30 and pcDNA3 ^1-p30-ROP2. Mice were injected with the recombinant plasmid to observe the immunoprotectivity of the nucleic acid vaccine by using ELISA for detection of total IgG and observing the survival time after tachyzoites challenge. Results The recombinant plasmids pcDNA3 ^1-p30 and pcDNA3 ^1-p30-ROP2 were constructed. Mice in pcDNA3 ^1-p30-ROP2 group showed higher IgG(P<0 ^05) and survived longer than those in pcDNA3 ^1-p30 group(P<0 ^01)after challenged with T.gondii. Conclusion Compound vaccine of genes from different stages of T.gondii elicits stronger immunoprotectivity in mice than a single gene vaccine.

关 键 词：PCDNA3 弓形虫 小鼠 表面抗原 基因疫苗 ROP 生存时间 真核细胞表达载体 重组表达质粒 基因组DNA 

分 类 号：R730.51[医药卫生—肿瘤] R780.2[医药卫生—临床医学]