| siRNA干扰lZP3 mRNA表达的研究 |
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| 作者姓名： | 庄淑珍 郭继娜 郑耀虎 李菁菁 张富春 |
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| 作者单位： | 新疆大学生命科学与技术学院分子生物学重点实验室 乌鲁木齐830046 |
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| 基金项目： | “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) |
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| 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。
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| 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 |
Study on the lZP3 mRNA Expression after siRNA Interference |
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| Authors: | Zhuang Shuzhen Guo Jin Zheng Yaohu Li Jingjing Zhang Fuchun |
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| Institution: | .Key Laboratory of Molecular Biology, College of Life Science and Technology, Xinjiang University, Urumqi 830046 |
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| Abstract: | 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. |
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| Keywords: | Lagurus lagurus RNA interference vector Zona pellucida 3 Hydrodynamics-based transfection method |
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