单 位：
新疆大学生命科学与技术学院分子生物学重点实验室
关键词：
草原兔尾鼠;RNA干扰载体;卵透明带3;尾静脉大容量快速注射
摘 要：
【目的】为探索新疆草原兔尾鼠(Lagurus lagurus)卵透明带3(lZP3)在精卵结合中的机理,有效的控制草原兔尾鼠的生育;【方法】将RNA干扰载体pGenesil-ZP31和真核表达质粒p-ACLC一起通过尾静脉大容量快速注射法(Hydrodynamics-based transfectionmethod,HD法)共注射小鼠。【结果】半定量RT-PCR和real-timePCR结果表明,2种质粒共注射时,pGenesil-ZP31的干涉作用至少发生在注射后的8h到10d之间;顺序性注射实验中,注射pGenesil-ZP31后间隔8h和1d再注射p-ACLC时,pGenesil-ZP31能在一定程度上干涉lZP3mRNA的表达,间隔3d以后lZP3mRNA的表达几乎不受影响,未出现干涉现象。而注射p-ACLC后间隔1h、2h、4h和8h再注射pGenesil-ZP31,lZP3mRNA的表达也受到了抑制,间隔24h时抑制现象则消失;【结论】以载体为基础的体内RNAi过程具有时间依赖性,为在卵母细胞水平进一步研究siRNA对lZP3的干涉机制提供借鉴。
译 名：
Study on the lZP3 mRNA Expression after siRNA Interference
作 者：
Zhuang Shuzhen, Guo Jina, Zheng Yaohu, Li Jingjing, Zhang Fuchun (Key Laboratory of Molecular Biology, College of Life Science and Technology, Xinjiang University, Urumqi 830046)
关键词：
Lagurus lagurus, RNA interference vector, Zona pellucida 3, Hydrodynamics-based transfection method
摘 要：
[Objective]In order to explore lZP3 gene in the oocytes and sperms combination mechanism and control the population of the Lagurus lagurus.[Method]The RNA interference vector pGenesil-ZP31 and the lZP3 expression vector were co-injected into the mice by Hydrodynamics-based transfection method (HD).[Results] Results of semi-quantative RT-PCR and real-time PCR showed that, the interference effect was occurred between 8 h and 10 d when co-injected the 2 vectors. Study of sequential injections revealed that prior injection of pGenesil-ZP31 produced a significant suppression at 8 h and 1 d, which disappeared after 3 days. However, injection of p-ACLC followed by pGenesil-ZP31 at 1 h, 2 h, 4 h and 8 h, the expression of lZP3 mRNA was also suppression, which disappeared after 24 hours.[Conclusion]Taken together, these results demonstrate that vector-based in vivo RNAi is a time-dependent process and offers the use for references in studying the mechanism of siRNA interference the Lagurus lagurus ZP3 at the oocytes level.
