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| 口蹄疫病毒3D蛋白在T4噬菌体衣壳表面的展示 | |
| 引用本文： | 覃健萍,徐维加,陈峰,马静云,谢青梅,姚兵华,曹永长,毕英佐.口蹄疫病毒3D蛋白在T4噬菌体衣壳表面的展示[J].农业生物技术学报,2008,16(2):190-195. |
| 作者姓名： | 覃健萍 徐维加 陈峰 马静云 谢青梅 姚兵华 曹永长 毕英佐 |
| 作者单位： | 1. 华南农业大学动物科学学院,广州,510640;广东温氏食品集团有限公司,新兴,527439 2. 云南出入境检验检疫局,昆明,650228 3. 华南农业大学动物科学学院,广州,510640 |
| 基金项目： | 广东省重大科技专项基金 |
| 摘 要： | 本文通过PCR技术获得3D基因，使用拼接重叠延伸聚合酶链反应对3D基因内部的EcoRⅠ位点进行定点突变，将突变后的3D基因克隆至原核表达质粒pSOC和T4噬菌体的穿梭质粒pR中，通过PCR及质粒酶切鉴定，分别获得阳性重组子pSOC-3D和pR-3D。将含有3D基因的重组质粒pSOC-3D质粒转化大肠杆菌BL21进行表达,SDS-PAGE检测表达产物。将含有3D基因的重组质粒pR-3D转化T4噬菌体的宿主菌E2，通过同源重组获得重组噬菌体T4-3D，经SDS-PAGE及Western- blot检测，证明展示在T4噬菌体表面的3D融合蛋白能与FMDV感染血清发生特异性反应。 |
| 关 键 词： | 口蹄疫病毒 非结构蛋白3D T4噬菌体展示 |
| 文章编号： | 1006-1304(2008)02-0190-06 |
| 收稿时间： | 2007-5-10 |
| 修稿时间： | 2007年5月10日 |
Display of Nonstructural Protein 3D of Foot-and-mouth disease virus on Capsid Surface of T4 Bacteriophage |
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| QIN Jian-ping,XU Wei-jia,CHEN Feng,MIA Jing-yun,XIE Qing-mei,YAO Bing-hua,CAO Yong-chang,BI Ying-zuo.Display of Nonstructural Protein 3D of Foot-and-mouth disease virus on Capsid Surface of T4 Bacteriophage[J].Journal of Agricultural Biotechnology,2008,16(2):190-195. | |
| Authors: | QIN Jian-ping XU Wei-jia CHEN Feng MIA Jing-yun XIE Qing-mei YAO Bing-hua CAO Yong-chang BI Ying-zuo |
| Abstract: | The nonstructural gene 3D of foot-and-mouth disease virus was amplified by polymerase chain reaction (PCR) from the recombinant plasmids T-3D. A point mutation was conducted at the EcoRⅠrestriction site of 3D gene by means of spliced ovelapping extension PCR. The mutated 3D gene was inserted into the pSOC expression plasmid and the T4 bacteriophage recombinant integration plasmids pR to obtain the recombinant plasmids pSOC-3D and pR-3D. The recombinant plasmids were selected and confirmed by PCR screening and EcoRⅠrestriction endonuclease digestion. The expression plasmid pSOC-3D was used to transform E. coli BL21 and the expression product was analyzed by SDS-PAGE. The recombinant integration plasmid pR-3D was transinfected E.coli E2 strain to obtain the recombinant phages T4-3D, and the protein bands could detected by SDS-PAGE and reacted specifically to serum from FMDV-infected animals. |
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