[Objective]The lentiviral-mediated PNX silencing vector was constructed and infected with GnRH secreting cells (GT1-7), and then screened and identified to obtain a stable PNX silencing cell line.[Method]A lentiviral silencing vector was constructed in conjunction with RNAi interference technology, and the virus was packaged in 293 T cells and assayed for titer.GnRH secreting cells (GT1-7) were infected, cells were screened by puromycin, the PNX silencing effect was detected by real-time PCR and western blot.[Result]The lentiviral-mediated PNX silencing vector was successfully constructed.The packaged lentivirus successfully infected GT1-7 cells, and the PNX silencing effect of PNX-silenced cell lines was confirmed by real-time PCR and western blot.[Conclusion]The lentivirus-mediated PNX silencing cell line was successfully constructed, which laid a foundation for further research on the role of PNX in animal reproductive development.%[目的]构建慢病毒介导的PNX沉默载体, 感染GnRH分泌细胞 (GT1-7) 后进行筛选鉴定, 获得稳定的PNX沉默细胞株.[方法]结合RNAi干扰技术构建出慢病毒沉默载体, 在293T细胞中包装病毒并测定滴度.感染GnRH分泌细胞 (GT1-7), 嘌呤霉素筛选得到稳定细胞后利用Real-time PCR和Western blot检测PNX沉默效果.[结果]慢病毒介导的PNX沉默载体构建成功, 包装的慢病毒成功感染GT1-7细胞, 并且Real-time PCR和Western blot证实了PNX沉默细胞株的PNX沉默效果.[结论]成功构建了慢病毒介导的PNX沉默细胞株, 为以后探索PNX对动物生殖发育的作用打下了基础.
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