| 猪流行性腹泻病毒LJB/03株的分离及膜蛋白基因的原核表达 |
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| 引用本文： | 吴凌,李一经.猪流行性腹泻病毒LJB/03株的分离及膜蛋白基因的原核表达[J].畜牧兽医学报,2005,36(10):1095-1099. |
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| 作者姓名： | 吴凌 李一经 |
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| 作者单位： | 1. 东北农业大学动物医学院,哈尔滨,158300;黑龙江八一农垦大学动物科技学院,大庆,163319
2. 东北农业大学动物医学院,哈尔滨,158300 |
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| 基金项目： | 黑龙江省科委“九五”攻关项目（G9988-1-1） |
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| 摘 要： | 猪流行性腹泻病毒（Porcine epidemic diarrhea virus，PEDV）和猪传染性胃肠炎病毒（Swine transmissible gastroenteritis virus，TGEV）同属于冠状病毒属的成员，主要引起2周龄仔猪以呕吐、腹泻、脱水为特征的一种急性病毒性传染病，死亡率达100％，已成为世界范围内猪的重要疫病之一。从病毒粒子形态、临床症状、病理变化和流行病学等方面无法区分两种病毒。国内外对两病的防制开展了大量的工作，但对于分子生物学诊断方面研究的较少。传统方法如电镜观察、病毒分离培养等不能有效地将TGEV与PEDV区分开，现代分子免疫学和分子生物学等的进展为新一代生物工程诊断试剂的研制开辟了新途径。
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| 关 键 词： | 猪流行性腹泻病毒 M基因 克隆 原核表达 免疫印迹 |
| 文章编号： | 0366-6964（2005）10-1095-05 |
| 收稿时间： | 2004-08-30 |
| 修稿时间： | 2004-08-30 |
Isolation of PEDV LJB/03 Strain and Expression of Its Membrane Protein in E. coli |
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| WU Ling,LI Yi-jing.Isolation of PEDV LJB/03 Strain and Expression of Its Membrane Protein in E. coli[J].Acta Veterinaria et Zootechnica Sinica,2005,36(10):1095-1099. |
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| Authors: | WU Ling LI Yi-jing |
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| Institution: | 1. College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China; 2. Animal Science and Technology College, Heilongjiang August First Land Reclamation University, Daqing 163319, China |
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| Abstract: | A strain of virus was isolated from faeces of diarrhea piglet using the Vero cells,and was identified as PEDV by RT-nested PCR,then was named as LJB/03.The 854 bp M gene of LJB/03 was amplified by RT-PCR,then was directly inserted into pMD18-T,and sequenced.The 854bp PM gene was sub-cloned into a prokaryotic expression vector pGEX-6P-1.A positive colony pGEX-6P-PM was translated into BL21(DE_3)plysS.High-level expression of recombinant fusion protein was obtained after induced by IPTG.43 ku fusion protein was tested by SDS-PAGE and can be recognized by the positive serum of PEDV by Western-blot.Through gel thin layer scanning analysis,the amount of target protein is over 33.7%.So the recombinant M protein can be a valuable diagnosis or therapeutic agent for the diseases. |
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| Keywords: | PEDV M gene cloning prokaryotic expression Western-blot |
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