猪圆环病毒2型ORF2基因的原核表达及反应原性分析
发布时间：2018-11-14 14:12
【摘要】：研究猪圆环病毒2型(PCV2)Cap蛋白的抗原性,为开发PCV2的检测方法奠定基础。以PCV2CAU0673毒株的DNA为模板,扩增获得702bp的目的片段,扩增产物克隆入pET30a(+)原核表达载体,构建pET30a-PCV2-ORF2重组质粒,转入大肠埃希菌BL21(DE3);在37℃以1mmol/L IPTG诱导表达6h;采用Ni-NTA树脂亲和层析纯化重组蛋白,并用不同浓度的尿素对纯化蛋白进行复性。SDSPAGE分析表明,该ORF2编码基因在大肠埃希菌中得到表达,蛋白大小约为34ku;Western blot检测结果表明,该重组Cap蛋白与PCV2阳性血清发生特异性反应,与NA-PRRSV和PPV1血清不发生交叉反应。成功构建了PCV2-ORF2原核表达载体,实现了在大肠埃希菌中的表达,纯化后的复性蛋白具有较好的反应原性,为猪圆环病毒2型检测方法建立或试剂盒的开发奠定了基础。
[Abstract]:The antigenicity of porcine circovirus type 2 (PCV2) Cap protein was studied. Using the DNA of PCV2CAU0673 strain as template, the target fragment of 702bp was amplified and cloned into pET30a () prokaryotic expression vector. The recombinant plasmid of pET30a-PCV2-ORF2 was constructed and transferred into BL21 (DE3) of Escherichia coli, and was induced by 1mmol/L IPTG for 6 h at 37 鈩，
本文编号：2331385
[Abstract]:The antigenicity of porcine circovirus type 2 (PCV2) Cap protein was studied. Using the DNA of PCV2CAU0673 strain as template, the target fragment of 702bp was amplified and cloned into pET30a () prokaryotic expression vector. The recombinant plasmid of pET30a-PCV2-ORF2 was constructed and transferred into BL21 (DE3) of Escherichia coli, and was induced by 1mmol/L IPTG for 6 h at 37 鈩，
本文编号：2331385
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