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中国农学通报

中国农学通报 ›› 2008, Vol. 24 ›› Issue (11): 15-18.doi: 10.11924/j.issn.1000-6850.2008-0462

所属专题： 生物技术； 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

猪圆环病毒Ⅱ型ORF2基因的克隆与表达

车勇良,庄向生,陈少莺,魏 宏,王隆柏,周伦江   

  • 收稿日期:2008-07-23 修回日期:2008-09-26 出版日期:2008-11-05 发布日期:2008-11-05

Cloning and Expression of ORF2 gene of Porcine Circovirus TypeⅡ

  • Received:2008-07-23 Revised:2008-09-26 Published:2008-11-05 Online:2008-11-05

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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。本研究用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4小时。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。

Abstract: 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.