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[目的]构建耐RNase酶的内含甲型H1N1流感病毒HA基因序列的病毒样颗粒。[方法]构建中间载体pET32a-MS2,将甲型H1N1流感病毒的HA基因片段连接中间载体上,构建原核表达载体pET32a-MS2-HA,转化宿主菌,诱导表达制备病毒样颗粒,对病毒样颗粒进行荧光定量RT-PCR检测和稳定性实验。[结果]表达载体经PCR、酶切鉴定分析后证实构建成功,荧光定量RT-PCR实验表明该病毒颗粒含有HA基因片段并且颗粒稳定性良好。[结论]成功构建了含甲型H1N1流感病毒HA基因序列的病毒样颗粒且稳定性良好,有望作为甲型H1N1流感病毒RNA检测的标准品和质控品。
[Objective] The aim of the study was to construct virus-like particles of RNase-resistant HA gene sequence of influenza A (H1N1) virus. [Method] The intermediate vector pET32a-MS2 was constructed and the HA gene fragment of influenza A (H1N1) virus was ligated into the intermediate vector. The prokaryotic expression vector pET32a-MS2-HA was constructed and transformed into host bacteria. The virus- Fluorescent quantitative RT-PCR detection and stability experiments. [Result] The expression vector was proved by PCR and restriction enzyme digestion analysis. The results of quantitative RT-PCR showed that the virus particle contained HA gene fragment and the particle stability was good. [Conclusion] The virus-like particles containing the HA gene sequence of influenza A (H1N1) virus were successfully constructed and were stable. It is expected to serve as a standard and quality control for the detection of influenza A (H1N1) virus RNA.