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Position: Home > Articles > Cloning and Expression of ORF2 Gene of Porcine Circovirus TypeⅡ Chinese Agricultural Science Bulletin 2008,24 (11) 15-18

猪圆环病毒Ⅱ型ORF2基因的克隆与表达

作  者:
车勇良;庄向生;陈少莺;魏宏;王隆柏;周伦江
单  位:
福建省农业科学院畜牧兽医研究所
关键词:
猪圆环病毒II型;ORF2;原核表达
摘  要:
猪圆环病毒II型(PCV2)是引起断奶后仔猪多系统衰竭综合症(PMWS)的重要病原。笔者用PCR方法扩增PCV2截短的ORF2基因并亚克隆至表达载体pGEX-6P-1,构建的重组表达质粒命名为pGEX-ORF2,将重组质粒转化大肠杆菌BL21(DE3),在37℃经IPTG诱导4h。PCV2的cap蛋白以包涵体形式得到有效表达。经SDS-PAGE及Western blot分析表明表达的重组蛋白分子量约为46KD,重组蛋白具有良好的免疫学活性。
译  名:
Cloning and Expression of ORF2 Gene of Porcine Circovirus TypeⅡ
作  者:
Che Yongliang1,2, Zhuang Xiangsheng1,2, Chen Shaoying1,2, Wei Hong1,2, Wang Longbai1,2, Zhou Lunjiang1,2(1Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture Sciences, Fuzhou Fujian 350013;2Fujian Animal Diseases Control Technology Development Center, Fuzhou Fujian 350013)
关键词:
porcine circovirus typeⅡ(PCV2), ORF2, prokaryotic expression
摘  要:
Porcine circovirus typeⅡ(PCV2) plays a crucial role in the pathogenesis of post-weaning multisystemic wasting syndrome(PMWS) in swine.The truncated ORF2 gene of PCV2 was amplified by PCR and the PCR product was cloned into plasmid PGEX-6P-1.The recombinant plasmid was name PGEX-ORF2.The PGEX-ORF2 was transformed into E.coli BL21(DE3) cells, and induced by IPTG at 37℃ for 4 hours.The recombinant PCV2 Cap protein was expressed efficiently in the form of inclusion body.SDS-PAGE and Western blotting analysis showed that the recombinant fusion protein had a molecular weight of approximate 46 kD, and had immunological reactivity.

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