siRNA干扰lZP3 mRNA表达的研究  

Study on the lZP3 mRNA Expression after siRNA Interference

作　　者：庄淑珍[1] 郭继娜[1] 郑耀虎[1] 李菁菁[1] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院分子生物学重点实验室,乌鲁木齐830046

出　　处：《中国农学通报》2007年第6期23-27,共5页Chinese Agricultural Science Bulletin

基　　金："国家自然科学基金资助项目"(No.30360062);"中国博士后科学基金资助项目"(2005038619);"新疆生物资源基因工程重点实验室开放课题资助项目"(XJDX0201-2006-03)

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。[Objective]In order to explore and control the population of the Lagurus IZP3 gene in the oocytes and sperms combination mechanism lagurus. [Method]The RNA interference vector pGenesil-ZP31 and the IZP3 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 IZP3 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.

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 

分 类 号：Q7[生物学—分子生物学]