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| 水稻黑条矮缩病病毒外壳蛋白基因S10的原核表达、多克隆抗体制备及应用 | |
| 引用本文： | 欧阳元龙,吴建祥,熊如意,周益军,周雪平. 水稻黑条矮缩病病毒外壳蛋白基因S10的原核表达、多克隆抗体制备及应用[J]. 中国水稻科学, 2010, 24(1): 25-30. DOI: 10.3969/j.issn.1001-7216.2010.01.05 |
| 作者姓名： | 欧阳元龙 吴建祥 熊如意 周益军 周雪平 |
| 作者单位： | 1. 南京师范大学,生命科学学院,江苏,南京,210046;浙江大学,农业与生物技术学院,生物技术研究所,浙江,杭州,310029;江苏省农业科学院,植物保护研究所,江苏,南京,210014 2. 浙江大学,农业与生物技术学院,生物技术研究所,浙江,杭州,310029 3. 江苏省农业科学院,植物保护研究所,江苏,南京,210014 |
| 基金项目： | 农业部公益性行业(农业)科研专项资助项目，现代农业产业技术体系建设专项资金资助项目，国家自然科学基金资助项目 |
| 摘 要： | 用RT-PCR方法从感染水稻黑条矮缩病毒(rice black-streaked dwarf virus,RBSDV)水稻中克隆该病毒的外壳蛋白基因S10,然后将此外壳蛋白基因再亚克隆到原核表达载体PET-32a中构建成重组原核表达载体pET32a-CP。将重组表达载体转化大肠杆菌BL21(DE3),经IPTG诱导,Ni+ NTA亲和柱纯化获得分子量约为76kD含硫氧还蛋白的融合蛋白。以纯化的重组蛋白为抗原免疫兔子制备RBSDV外壳蛋白的多克隆抗体,并用制备的多克隆抗体建立了可靠、灵敏、特异的检测RBSDV的免疫捕获RT-PCR及Dot-blot ELISA方法,为该水稻病毒病的诊断提供技术支持。 |
| 关 键 词： | 水稻黑条矮缩病病毒 原核表达 多克隆抗体 免疫捕获反转录聚合酶链式反应 斑点酶联免疫吸附测定 |
| 收稿时间： | 1900-01-01; |
Prokaryotic Expression of Coat Protein Gene S10 of Rice Black-Streaked Dwarf Virus,and Preparation and Application of Its Polyclonal Antibody |
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| OUYANG Yuan-long,WU Jian-xiang,XIONG Ru-yi,ZHOU Yi-jun,ZHOU Xue-ping. Prokaryotic Expression of Coat Protein Gene S10 of Rice Black-Streaked Dwarf Virus,and Preparation and Application of Its Polyclonal Antibody[J]. Chinese Journal of Rice Science, 2010, 24(1): 25-30. DOI: 10.3969/j.issn.1001-7216.2010.01.05 | |
| Authors: | OUYANG Yuan-long WU Jian-xiang XIONG Ru-yi ZHOU Yi-jun ZHOU Xue-ping |
| Affiliation: | OUYANG Yuan-long1,2,3,WU Jian-xiang2,XIONG Ru-yi3,ZHOU Yi-jun3,ZHOU Xue-ping2(1College of Life Science,Nanjing Normal University,Nanjing 210046,China,2Institute of Biotechnology,College of Agriculture & Biotechnology,Zhejiang University,Hangzhou 310029,3Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China) |
| Abstract: | The full length cDNA of rice black-streaked dwarf virus (RBSDV) segment 10 (S10) which encoded coat protein was cloned from the virus infected rice samples by RT-PCR,and subcloned into a prokaryotic expression vector pET-32a. The recombinant prokaryotic expression vector (pET-32a-CP) was used to transform Escherichia coli BL21 (DE3). A 76 kD TrxA fusion protein was obtained with induction of IPTG and purification of Ni+ NTA affinity column. The purified recombinant protein was used to immunize rabbits for p... |
| Keywords: | rice black-streaked dwarf virus prokaryotic expression polyclonal antibody immunocapture RT-PCR dot-blot ELISA |
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