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中国农学通报

中国农学通报 ›› 2007, Vol. 23 ›› Issue (6): 23-23.

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

siRNA干扰lZP3 mRNA表达的研究

庄淑珍,郭继娜,郑耀虎,李菁菁,张富春   

  • 出版日期:2007-06-05 发布日期:2007-06-05

Study on the lZP3 mRNA Expression after siRNA Interference

Zhuang Shuzhen, Guo Jina, Zheng Yaohu, Li Jingjing, Zhang Fuchun   

  • Online:2007-06-05 Published:2007-06-05

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 transfection method, HD法）共注射小鼠。【结果】半定量RT-PCR和real-time PCR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8 h到10 d之间；顺序性注射实验中，注射pGenesil-ZP31后间隔8 h和1 d再注射p-ACLC时，pGenesil-ZP31能在一定程度上干涉lZP3 mRNA的表达，间隔3 d以后lZP3 mRNA的表达几乎不受影响，未出现干涉现象。而注射p-ACLC后间隔1 h、2 h、4 h和8 h再注射pGenesil-ZP31，lZP3 mRNA的表达也受到了抑制，间隔24 h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。

关键词: 野生蔬菜, 野生蔬菜, 研究与开发, 学科, 科学价值

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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