通过基因克隆和表达产生的针对桃拉综合征病毒（TSV）VP1和VP3衣壳蛋白的多克隆抗体。

Polyclonal antibodies specific for VP1 and VP3 capsid proteins of Taura syndrome virus (TSV) produced via gene cloning and expression.

作者信息

Chaivisuthangkura Parin, Tejangkura Thanawan, Rukpratanporn Sombat, Longyant Siwaporn, Sithigorngul Weerawan, Sithigorngul Paisarn

机构信息

Department of Biology, Faculty of Science, Srinakharinwirot University, Bangkok, Thailand.

出版信息

Dis Aquat Organ. 2006 Apr 6;69(2-3):249-53. doi: 10.3354/dao069249.

DOI:10.3354/dao069249

PMID:16724569

Abstract

Capsid protein genes VP1 and VP3 of Taura syndrome virus (TSV) were cloned into pGEX-6P-1 expression vector and transformed into Escherichia coli BL21. After induction, recombinant VP1 (rVP1) and recombinant VP3 (rVP3) were produced, purified by SDS-PAGE and used for immunization of Swiss mice for antisera production. Anti-rVP1 and anti-rVP3 antisera showed specific immunoreactivities to rVP1 and rVP3 proteins, respectively, by Western blot assay and also yielded good results for detection of TSV in various shrimp tissues by immunohistochemistry. This is the first step towards our target of preparing monoclonal antibodies specific to rVP1 and rVP3 for use in simple immuno-diagnostic test kits for TSV detection and identification.

摘要

将桃拉综合征病毒（TSV）的衣壳蛋白基因VP1和VP3克隆到pGEX-6P-1表达载体中，并转化到大肠杆菌BL21中。诱导后，产生重组VP1（rVP1）和重组VP3（rVP3），通过SDS-PAGE进行纯化，并用于免疫瑞士小鼠以产生抗血清。通过蛋白质免疫印迹分析，抗rVP1和抗rVP3抗血清分别对rVP1和rVP3蛋白表现出特异性免疫反应，并且在通过免疫组织化学检测各种虾组织中的TSV时也取得了良好的结果。这是我们制备针对rVP1和rVP3的单克隆抗体以用于TSV检测和鉴定的简单免疫诊断试剂盒这一目标的第一步。