Key Laboratory of Molecular Animal Nutrition, Ministry of Education, Feed Science Institute, Zhejiang University, Hangzhou 310029, China.
Virol Sin. 2011 Aug;26(4):260-6. doi: 10.1007/s12250-011-3202-0. Epub 2011 Aug 17.
The gene encoding the VP28 envelope protein of White spot syndrome virus (WSSV) was cloned into expression vector pET-30a and transformed into the Escherichia coli strain BL21. After induction, the recombinant VP28 (rVP28) protein was purified and then used to immunize Balb/c mice for monoclonal antibody (MAb) production. It was observed by immuno-electron microscopy the MAbs specific to rVP28 could recognize native VP28 target epitopes of WSSV and dot-blot analysis was used to detect natural WSSV infection. Competitive PCR showed that the viral level was approximately 10(4) copies/mg tissue in the dilution of gill homogenate of WSSV-infected crayfish at the detection limit of dot-blot assay. Our results suggest that dot-blot analysis with anti-rVP28 MAb could rapidly and sensitively detect WSSV at the early stages of WSSV infection.
白斑综合征病毒（WSSV）VP28 囊膜蛋白的编码基因被克隆到表达载体 pET-30a 中，并转化到大肠杆菌菌株 BL21 中。诱导后，重组 VP28（rVP28）蛋白被纯化，然后用于免疫 Balb/c 小鼠以产生单克隆抗体（MAb）。通过免疫电子显微镜观察到，针对 rVP28 的 MAb 能够识别 WSSV 的天然 VP28 靶表位，并且点印迹分析用于检测天然 WSSV 感染。竞争性 PCR 显示，在斑点印迹分析的检测限下，WSSV 感染螯虾鳃匀浆的稀释度中病毒水平约为 10(4) 拷贝/mg 组织。我们的结果表明，用抗-rVP28 MAb 进行点印迹分析可以在 WSSV 感染的早期快速和敏感地检测到 WSSV。
