| 对虾白斑综合征病毒囊膜蛋白VP53B原核表达及抗血清的制备 |
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| 引用本文： | 姚迁会,覃冰,潘迎捷,王永杰.对虾白斑综合征病毒囊膜蛋白VP53B原核表达及抗血清的制备[J].微生物学通报,2015,42(10):1977-1983. |
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| 作者姓名： | 姚迁会 覃冰 潘迎捷 王永杰 |
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| 作者单位： | 1. College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China;2. Laboratory of Quality ＆ Safety Risk Assessment for Aquatic Products on Storage and Preservation, Ministry of Agriculture, Shanghai 201306, China,1. College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China;2. Laboratory of Quality ＆ Safety Risk Assessment for Aquatic Products on Storage and Preservation, Ministry of Agriculture, Shanghai 201306, China,1. College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China;2. Laboratory of Quality ＆ Safety Risk Assessment for Aquatic Products on Storage and Preservation, Ministry of Agriculture, Shanghai 201306, China and 1. College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China;2. Laboratory of Quality ＆ Safety Risk Assessment for Aquatic Products on Storage and Preservation, Ministry of Agriculture, Shanghai 201306, China |
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| 基金项目： | 教育部高等学校博士点基金项目(No. 20133104110006)；上海市教委科研创新重点项目(No. 14ZZ144) |
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| 摘 要： | 【目的】研究对虾白斑综合征病毒(White spot syndrome virus，WSSV)囊膜蛋白sVP53B克隆、表达、纯化及抗血清制备。【方法】根据WSSV囊膜蛋白基因序列，设计引物，PCR扩增出功能序列(Svp53B)，构建到pET-16b载体后，转化至大肠杆菌Rosetta 2诱导表达，用SDS-PAGE、Western blotting检测优化表达。表达产物采用Ni-NTA琼脂糖磁珠进行纯化、割胶回收融合蛋白，以纯化的Svp53B-his为抗原，免疫兔子获得多克隆抗体，通过间接ELISA检测抗体的效价。【结果】构建重组质粒pET-16b-Svp53B，在大肠杆菌Rosetta 2中以1 mmol/L IPTG诱导表达量最高，主要以包涵体形式表达。纯化包涵体蛋白免疫兔子，获得多克隆血清，效价达到1:150 000。【结论】原核表达并纯化得到高纯度的WSSV囊膜蛋白sVP53B，制备的兔源多克隆血清亲和力高、特异性好，这对后期进一步研究VP53B与经口侵染相关功能奠定了基础。
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| 关 键 词： | 对虾白斑综合征病毒，囊膜蛋白，VP53B，原核表达，多克隆血清 |
Prokaryotic expression and serum preparation of envelope protein VP53B of white spot syndrome virus (WSSV) |
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| YAO Qian-Hui,QIN Bing,PAN Ying-Jie and WANG Yong-Jie.Prokaryotic expression and serum preparation of envelope protein VP53B of white spot syndrome virus (WSSV)[J].Microbiology,2015,42(10):1977-1983. |
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| Authors: | YAO Qian-Hui QIN Bing PAN Ying-Jie and WANG Yong-Jie |
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| Abstract: | |
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| Keywords: | White spot syndrome virus Envelope protein VP53B Prokaryotic expression Polyclonal serum |
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