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目的RT-PCR扩增、克隆SARS冠状病毒N蛋白基因。方法根据GenBank数据库中TOR2株的全基因组序列,利用Primer Premier 5.0软件设计引物RT-PCR巢式扩增SARS冠状病毒的N基因,PCR产物克隆后进行测序鉴定。结果序列分析表明,pMD18-T载体中已成功重组了N基因。结论N蛋白基因的扩增、克隆成功,为N蛋白的表达、N蛋白结构与功能的研究奠定了基础。
Objective To amplify and clone SARS-CoV N protein by RT-PCR. Methods The N gene of SARS coronavirus was amplified by RT-PCR using primers designed by Primer Premier 5.0 according to the complete genome sequence of TOR2 strain in GenBank. The PCR products were cloned and sequenced. Results Sequence analysis showed that N gene was successfully recombined in pMD18-T vector. Conclusion The amplification and cloning of N protein gene is successful, which lays a foundation for the study of N protein expression and N protein structure and function.