猪圆环病毒Ⅱ型ORF2基因的克隆与表达

Cloning and Expression of ORF2 Gene of Porcine Circovirus TypeⅡ

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。

Porcine circovirus type Ⅱ （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.