截短表达的TTSuV2 ORF1重组蛋白对猪血清的免疫学反应

School of Life Science，Foshan University，Foshan 528000，Guangdong Province，China

摘要:

摘要:【目的】本文旨在了解猪细环病毒2(Torque teno sus virus 2，TTSuV2)病毒蛋白对猪血清的免疫学反应，明确TTSuV2 ORF1编码的氨基酸序列上特定片段的免疫原性，为TTSuV2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TTSuV2毒株为研究对象，在大肠杆菌表达系统中克隆表达了TTSuV2ORF1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(TTSuV2 ORF1a与TTSuV2 ORF1ab)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TTSuV2 ORF1a与TTSuV2 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TTSuV2 ORF1a与含有416个氨基酸的重组TTSuV2 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】TTSuV2 ORF1a与TTSuV2 ORF1ab相互重叠的179个氨基酸序列上(168－346)含有TTSuV2 ORF1关键的B细胞表位，重组蛋白TTSuV2 ORF1a适用于TTSuV2血清抗体免疫学检测。

Abstract:

Abstract:［Objective］We studied the serological response between the special regions on the Torque teno sus virus 2 (TTSuV2) ORF1 coded protein and the porcine sera from conventional pigs.［Methods］Based on a Chinese TTSuV2 strain from Guangdong province，two overlapped virus proteins were expressed from Escherichia coli．Then，purified recombinant TTSuV2 ORF1a and TTSuV2 ORF1ab proteins were used as the antigens in the Western Blotting and ELISA assay.［Results］The recombinant TTSuV2 ORF1a and TTSuV2 ORF1ab 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 TTSuV2 ORF1a and TTSuV2 ORF1ab proteins antigenic assay．The results of the following Western Blotting assay indicated that the TTSuV2-specific IgG antibodies were contained in pig sera． ［Conclusion］The truncated TTSuV2 ORF1a protein (positions 168 to 346 corresponding to TTSuV2 GDIMA1) contains important B cell epitopes which can stimulate immune system antibody secretion．The truncated TTSuV2 ORF1a protein could be effective in TTSuV2 immunodiagnosis.