果蝇钠通道DmNav22在双氧木脂素A作用机理研究中的应用  

Application of DrosophilaSodium Channel DmNav22 in Investigating the Action Mode of Haedoxan A

作　　者：闫秀芳[1,2] 郄杏桃 吴文君[1,2] 胡兆农[1,2] 

机构地区：[1]西北农林科技大学植物保护学院农药研究所,陕西杨凌712100 [2]西北农林科技大学陕西省植物源农药研究与开发重点实验室,陕西杨凌712100

出　　处：《西北农业学报》2017年第4期635-640,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(31672055);陕西省农业科技创新与攻关(2015NY033)~~

摘　　要：建立以爪蟾卵母细胞为载体的果蝇钠通道表达系统,研究双氧木脂素A的杀虫分子机理。将果蝇钠通道DmNav22与TipE亚基基因分别进行体外转录,获得DmNav22与TipE亚基基因的cRNA,将DmNav22与TipE的cRNA等质量混合,显微注射至新鲜的非洲爪蟾卵母细胞,在ND96溶液、18℃培养12~18h后采用双电极电压钳技术记录其电流,测定双氧木脂素A对钠电流的影响。结果表明,注射cRNA的非洲爪蟾卵母细胞,可记录到明显的果蝇钠通道电压门控内向电流,未注射的对照组细胞则未记录到钠电流;双氧木脂素A使钠通道失活电压依赖性向超极化方向移动。说明,果蝇钠通道在非洲爪蟾卵母细胞中成功表达,为进一步研究双氧木脂素A的作用机理奠定基础。To investigate the molecular mechanism of haedoxan A （HA） against insect pests,gene expression system of Drosophila sodium channel in Xenopus laevis oocytes was established, cRNAs of Drosophila sodium channel subunits DmNav22 and TipE were obtained by transcription in vitro, which were mixed with equal amount of substance and then microinjected in the fresh X. laevis oocytes. The oocytes were incubated in ND96 buffer at 18 ℃ for 12--18 hours after microinjection and the sodium channel current was detected by two-electrode voltage clamp （TEVC）. The results showed that the voltage-gated inward current was recorded in the oocytes injected cRNA,while no current was recorded in the blank oocytes. HA shifted the voltage dependence of Drosophila sodium channels inactivation to the hyperpolarizing direction. These results demonstrated that the Drosophila sodium channel was expressed in X. laevis oocytes successfully and can be used to further study the insecticidal mechanism of HA.

关 键 词：果蝇钠通道 双电极电压钳 非洲爪蟾卵母细胞 双氧木脂素A 

分 类 号：S481[农业科学—农药学]