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目的克隆化非结构蛋白基因NS5A。方法根据NCBI的Genbank上公布的HCV-1b型基因序列,设计PCR产物为NS5A的特异性引物,以HCV-1b型RNA为模板,利用RT-PCR方法扩增出1341 bp片段,连接至pGEM-T载体并送测序,测序结果表明NS5A已克隆成功。结果成功克隆了丙肝病毒非结构蛋白基因NS5A。结论NS5A是一种典型的病毒基因组编码的多功能蛋白,NS5A的克隆化为进一步研究NS5A的分子生物学机制和探索新型治疗技术奠定基础。
Objective To clone the nonstructural protein gene NS5A. Methods According to the sequence of HCV-1b gene published on the GenBank of NCBI, a PCR product was designed as a specific primer for NS5A. A 1341 bp fragment was amplified by RT-PCR using HCV-1b RNA as a template and ligated to pGEM- T vector and send sequencing, sequencing results show that NS5A has been cloned successfully. Results The hepatitis C virus nonstructural protein gene NS5A was successfully cloned. Conclusion NS5A is a typical multifunctional protein encoded by the viral genome. The cloning of NS5A lays the foundation for further research on the molecular mechanism of NS5A and exploration of new therapeutic techniques.