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当前位置: 首页 > 文章 > 慢病毒介导PNX沉默细胞株的构建与鉴定 安徽农业科学 2019 (2) 94-95+110
Position: Home > Articles > Construction and Identification of Lentivirus-mediated PNX Silencing Cell Line Journal of Anhui Agricultural Sciences 2019 (2) 94-95+110

慢病毒介导PNX沉默细胞株的构建与鉴定

作  者：
丁毅飞;曹春雨;王凭青
单  位：
重庆大学生物工程学院
关键词：
慢病毒;PNX;生殖
摘  要：
[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。
译  名：
Construction and Identification of Lentivirus-mediated PNX Silencing Cell Line
作  者：
DING Yi-fei;CAO Chun-yu;WANG Ping-qing;Bioengineering Institute of Chongqing University;
单  位：
DING Yi-fei%CAO Chun-yu%WANG Ping-qing%Bioengineering Institute of Chongqing University
关键词：
Lentivirus;;PNX;;Reproduction
摘  要：
[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.

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