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[目的]探讨Dorsal与葱蝇滞育发育的关系。[方法]以葱蝇(Delia antiqua)为试虫,采用RACE方法克隆了葱蝇Dorsal的全长cDNA序列;将该序列推导的氨基酸序列在GenBank搜寻同源序列,与搜寻到的14个代表性昆虫Dorsal序列进行相似性比较,并进行系统进化分析;采用半定量RT-PCR方法分析了Dorsal基因在葱蝇非滞育、夏滞育、冬滞育蛹中的表达情况。[结果]克隆了葱蝇Dorsal基因的cDNA序列,全长2 412 bp,开放阅读框1 974 bp,编码657个氨基酸,等电点PI约8.5,分子量72.9 kDa。与GenBank中的其他14个昆虫代表性Dorsal序列相似性比较和进化分析显示葱蝇的Dorsal与黑腹果蝇(Drosophila melanogaster)、拟暗果蝇(Drosophlia pseudoob-scura)的Dorsal序列相似性最大。半定量分析显示葱蝇Dorsal基因在葱蝇冬滞育、夏滞育、非滞育蛹滞育发育关键期的表达量均明显升高,尤其在滞育后期表达量最高,初步判断Dorsal基因与葱蝇滞育发育相关。[结论]为进一步开展Dorsal基因在昆虫滞育发育过程中的功能研究奠定了基础。
[Objective] The research aimed to discuss the relationship between Dorsal and diapause diapause development. [Method] The full-length cDNA sequence of Drosophila line Dorsal was cloned by RACE from Delia antiqua. The amino acid sequence deduced from this sequence was searched for homologous sequences in GenBank and compared with 14 representative Insect Dorsal sequences were compared and analyzed phylogenetically. The expression of Dorsal gene in non-diapause, summer diapause, winter diapause pupae of onion fly was analyzed by semi-quantitative RT-PCR. [Result] The cDNA sequence of Dorsal was cloned. The full - length cDNA was 2 412 bp in length and 1 974 bp in open reading frame, encoding 657 amino acids. The isoelectric point PI was 8.5 and the molecular weight was 72.9 kDa. The Dorsal sequence similarity analysis and phylogenetic analysis with other 14 representative insects in GenBank showed that Dorsal sequence similarity of Dorsal from Drosophila melanogaster and Drosophila pseudoob-scura was the highest. Semi-quantitative analysis showed that the expression level of Dorsal was significantly increased during the diapause stage of diapause, diapause, diapause and non-diapause stages, especially in the late diapause stage. The Dorsal gene and Diaofa diapause developmental related. [Conclusion] This laid the foundation for further study on the function of Dorsal gene during the development of insect diapause.