| O型口蹄疫病毒VP1基因的高效可溶表达及抗原性分析 |
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| 引用本文： | 谢青梅,李少璃,陈丽,覃健萍,马静云,毕英佐,曹永长.O型口蹄疫病毒VP1基因的高效可溶表达及抗原性分析[J].农业生物技术学报,2006,14(4):526-529. |
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| 作者姓名： | 谢青梅 李少璃 陈丽 覃健萍 马静云 毕英佐 曹永长 |
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| 作者单位： | 1. 华南农业大学动物科学学院,广州,510642
2. 中山大学生命科学学院 |
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| 基金项目： | 广东省重大科技专项基金 |
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| 摘 要： | 将猪源Ｏ型口蹄疫病毒(Foot-and-mouth disease virus, FMDV)VP1基因克隆到原核表达载体pMBX上，成功地构建重组表达质粒pMBX-VP1。将其转化大肠杆菌(Escherichia coli)BL21(DE3)感受态细胞中，经SDS-PAGE分析，融合蛋白分子量约为58 kD，表达产物占菌体总蛋白的35.5％。 经蛋白可溶性分析，目的蛋白在裂解沉淀中占6.8％，在裂解上清中占31.2％，融合蛋白绝大部分是以可溶形式表达的。经Western blot证实，可溶表达的融合蛋白与FMDV阳性血清具有良好的免疫反应性。将可溶表达的融合蛋白用50%Ni-NTA树脂纯化，用融合蛋白作包被抗原，通过ELISA方法，能特异性地检测出口蹄疫阳性血清。
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| 关 键 词： | 口蹄疫病毒(FMDV) VP1基因 高效可溶表达 抗原性 |
| 文章编号： | 1006-1304（2006）04-0526-04 |
| 收稿时间： | 2005-9-13 |
| 修稿时间： | 2006-3-30 |
High Soluble Expression and Antigenicity of VP1 Gene of Foot-and-mouth disease virus |
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| XIE Qing-mei,LI Shao-li,CHEN Li,QIN Jian-ping,MA Jing-yun,BI Ying-zuo,CAO Yong-chang.High Soluble Expression and Antigenicity of VP1 Gene of Foot-and-mouth disease virus[J].Journal of Agricultural Biotechnology,2006,14(4):526-529. |
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| Authors: | XIE Qing-mei LI Shao-li CHEN Li QIN Jian-ping MA Jing-yun BI Ying-zuo CAO Yong-chang |
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| Institution: | College of Animal Science, South China Agricultural University, Guangzhou 510640, China |
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| Abstract: | Foot-and-mouth disease virus (FMDV) VP1 gene was amplified by RT-PCR. The VP1 fragment was inserted into plasmid pMBX to obtain recombinant plasmid pMBX-VP1. The pMBX-VP1 was transformed into Escherichia coli BL-21(DE3) to induce VP1 gene fusion expression with 1 mmol/L Isopropylthio-β-D-galactoside (IPTG). The fusion protein band with molecular weight of 58 kD was visible on the SDS-PAGE gel. The density scanning showed that the largest amount of the fusion protein was 35.5% of total bacterial protein. The amounts of the soluble form of the fusion in supernatant of lysate was up to 31.2%, and 6.8% in sediment. Western blot result shawed that the expression products could specifically reacted with the antisera against FMDV. The soluble fusion protein was purified by 50%Ni-NTA affinity chromatography and used as an antigen, and FMDV antibody could be detected by ELISA method. |
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| Keywords: | Foot and mouth disease virus ( FMDV) VP1 gene high soluble expression antigenicity |
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