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河北地区猪圆环病毒二型ORF2基因片段的克隆与原核表达

DOI: 10.7668/hbnxb.2008.04.012, PP. 51-54

Keywords: 猪圆环病毒二型,ORF2基因片段,克隆,原核表达

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Abstract:

以PCV2PK-15细胞毒提取的DNA为模板,用PCR扩增PCV2ORF2基因的后部约600bp片段;对PCR产物回收纯化后与载体pMD19-T进行连接、转化,经酶切和序列分析后亚克隆到原核表达载体pET-32a中,构建重组表达质粒pET-32a-ORF2,转入大肠埃希氏菌BL21(DE3)中,以IPTG诱导表达,进行SDS-PAGE电泳和Western-Blotting分析。结果表明,重组表达质粒在大肠中所表达的融合蛋白相对分子量为40kDa,Western-blotting分析表明该蛋白具有PCV2抗原性。

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