柯萨奇B4病毒非结构蛋白P_2C重组质粒的构建

Construction of recombinant nonstructural P_2C plasmid in coxsackie B4 virus

目的:构建柯萨奇B4病毒非结构蛋白P2C重组质粒。方法:用RT-PCR技术,从柯萨奇B4病毒中钓取非结构蛋白P2CcDNA片段,克隆入PUCm-T载体,转化大肠杆菌DH5α,构建重组质粒。结果:以柯萨奇B4病毒的总RNA为模板,扩增出987bp大小的片段,克隆入PUCm-T载体,用BamHⅠ、HindⅢ双酶切鉴定,与非结构蛋白P2C的PCR扩增产物片段大小一致。结论:成功地构建了柯萨奇B4病毒非结构蛋白P2C重组质粒。

Objective. To consttruct the recombinant nonstructural P2C plasmid in coxsackie B4 virus. Methods：Using the RT-PCR, we fishing the nonstructural protein P2C cDNA fraction from Coxsackie B4 virus（CVB4） and cloning into the PUCm-T vector, then transfer it into E. coli to construct the recombinant plasmid. Results： Using the total RNA of Coxsackie B4 virus as model, we amplied 987 bp fraction and cloned into PUCm-T vector identifying with BamH Ⅰand Hind Ⅲ double enzyme. Its products were consistent with nonstructural protein P22 CPCR productions. Conclusion： The recombinant nonstructural P2C plasmid was constructed succesfully in coxsackie B4 virus.