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Fe-S蛋白(rieske iron-sulfur protein,RISP)是线粒体复合物Ⅲ的关键蛋白亚基之一,在呼吸链电子传递中起着重要作用。本文利用RT-PCR技术和RACE方法获得了斜纹夜蛾Spodoptera litura(Fab.)线粒体复合物ⅢRISP基因,该基因序列全长4413bp,含有3个内含子;开放阅读框长816bp,编码271个氨基酸。经氨基酸序列比对,与鳞翅目家蚕Bombyx mori和小菜蛾Plutella xylostella的Fe-S蛋白一致性较高,分别为83%和79%,与其他目昆虫的RISP一致性较低。qRT-PCR分析结果表明,SlitRISP基因在斜纹夜蛾不同发育阶段的表达水平存在显著差异,即幼虫期和成虫期的表达量显著高于卵期和蛹期,其中幼虫期又以第4和5龄幼虫表达水平显著高于初孵幼虫以及第1,2,3和6龄幼虫。斜纹夜蛾SlitRISP基因的克隆成功为今后对其功能研究以及作为靶标设计新型高效杀虫剂提供了基础。
Fe-S protein (rps-iron-sulfur protein, RISP) is one of the key protein subunits of mitochondrial complex III, which plays an important role in respiratory chain electron transfer. In this study, the mitochondrial ⅢRISP gene of Spodoptera litura (Fab.) Was obtained by RT-PCR and RACE. The full length of this gene is 4413bp and contains 3 introns. The open reading frame is 816bp in length and encodes 271 amino acids . The results of amino acid sequence alignment showed that there was a high identity of 83% and 79% to the Fe-S protein of Bombyx mori and Plutella xylostella, respectively, which was consistent with other species of insects. The results of qRT-PCR analysis showed that the expression level of SlitRISP gene at different developmental stages of Spodoptera litura was significantly different, that is, the expression level of SlitRISP gene in larval stage and adult stage was significantly higher than that in egg stage and pupal stage. The larval expression level was significantly higher than the newly hatched larvae as well as the 1st, 2nd, 3rd and 6th larvae. The successful cloning of the SlitRISP gene from Spodoptera litura provides the basis for its future functional studies and the design of new and highly effective insecticides as targets.