截短表达的TTSuV2 ORF1重组蛋白对猪血清的免疫学反应  

Serological tests of the truncated TTSuV2 ORF1 recombinant proteins with the porcine sera

作　　者：李中圣 陈轶雯[1,2] 白挨泉[3] 陈善真 王璐璐 姚火春[2] 王贵平 

机构地区：[1]广东海大畜牧兽医研究院有限公司,广东广州511400 [2]南京农业大学动物医学院,江苏南京210095 [3]佛山科学技术学院生命科学学院,广东佛山528000

出　　处：《微生物学报》2015年第2期227-234,共8页Acta Microbiologica Sinica

基　　金：广东省科技计划项目(2012A020602072)~~

摘　　要：【目的】本文旨在了解猪细环病毒2(Torque teno sus virus 2,TTSu V2)病毒蛋白对猪血清的免疫学反应,明确TTSu V2 ORF1编码的氨基酸序列上特定片段的免疫原性,为TTSu V2的免疫学检测方法建立提供实验依据。【方法】以本实验室测序的TTSu V2毒株为研究对象,在大肠杆菌表达系统中克隆表达了TTSu V2ORF1基因上两相互重叠的基因片段,对表达产物进行纯化,分析了两重组蛋白(TTSu V2 ORF1a与TTSu V2ORF1ab)对猪血清抗体的免疫学反应。【结果】应用标签抗体进行Western Blotting分析的结果显示,重组TTSu V2 ORF1a与TTSu V2 ORF1ab蛋白成功表达。应用建立的ELISA方法对212份猪血清进行检测的结果表明,含有179个氨基酸的重组TTSu V2 ORF1a与含有416个氨基酸的重组TTSu V2 ORF1ab蛋白之间有显著的相关性,二者对猪血清的反应基本一致。采用猪血清进行Western Blotting分析的结果显示,血清抗体可特异性识别两重组蛋白。【讨论】TTSu V2 ORF1a与TTSu V2 ORF1ab相互重叠的179个氨基酸序列上(168-346)含有TTSu V2 ORF1关键的B细胞表位,重组蛋白TTSu V2 ORF1a适用于TTSu V2血清抗体免疫学检测。[Objective] We studied the serological response between the special regions on the Torque teno sus virus 2（ TTSu V2） ORF1 coded protein and the porcine sera from conventional pigs. [Methods]Based on a Chinese TTSu V2 strain from Guangdong province,two overlapped virus proteins were expressed from Escherichia coli. Then,purified recombinant TTSu V2 ORF1 a and TTSu V2 ORF1 ab proteins were used as the antigens in the Western Blotting and ELISA assay. [Results] The recombinant TTSu V2 ORF1 a and TTSu V2 ORF1 ab proteins were identified with the special tag monoclonal antibody. The results of the ELISA tests shown that there were significant relationships between two groups of dates from the recombinant TTSu V2 ORF1 a and TTSu V2 ORF1 ab proteins antigenic assay. The results of the following Western Blotting assay indicated that the TTSu V2-specific Ig G antibodies were contained in pig sera. [Conclusion] The truncated TTSu V2 ORF1 a protein（ positions 168 to 346 corresponding to TTSu V2 GDIMA1） contains important B cell epitopes which can stimulate immune system antibody secretion. The truncated TTSu V2 ORF1 a protein could be effective in TTSu V2 immunodiagnosis.

关 键 词：猪细环病毒2 ORF1蛋白 表达和纯化 免疫学分析 

分 类 号：S852.65[农业科学—基础兽医学]