家蚕表皮型几丁质合成酶基因BmCHSA-2a的鉴定及对激素响应

Identification of Epidermal Chitin Synthaes BmCHSA-2a in Bombyx mori and Its Response to Hormones

  • 摘要: 文中采用5'Race方法获得了几丁质合成酶基因(BmCHSA-2a)的第1外显子(Exon 1, 141 bp)和第2外显子(Exon 2a,106 bp),证明外显子2a在鳞翅目昆虫几丁质合成酶中高度保守. 根据该外显子序列设计引物,分析BmCHSA-2a对激素响应分析. 结果表明蜕皮激素(Ecdysone ,20E)抑制BmCHSA-2a的表达,而保幼激素(Juvenile Hormone, JH)间接促进其表达. 根据家蚕基因组所预测的启动子序列,扩增了调控BmCHSA-2a表达的启动子,证明蜕皮激素抑制启动子活性,并存在大量蜕皮激素通路BmFTZ-F1的调控元件,对BmFTZ-F1表达调控的进一步研究表明,蜕皮激素抑制其表达. 研究结果初步分析了家蚕表皮几丁质合成酶BmCHSA-2a可能的表达机制,为深入研究其表达调控机理及发现害虫防治新靶标提供了理论基础.

     

    Abstract: In this study, exon 1 (141 bp) and exon 2a (106 bp) of BmCHSA-2a were cloned by using 5'Race, and the mRNA levels of BmCHSA-2a in response to hormones were analyzed by using the primers based on the sequence of exon 2a, which is conservative in insects. The results showed that the expression of BmCHSA-2a was inhibited by ecdysone (20E), but was indirectly induced by juvenile hormone (JH). Promoter activity determination confirmed that the transcription of BmCHSA-2a was inhibited. Further analysis revealed that a lot of BmFTZ-F1 cis-regulation elements existed in the promoter sequence of BmCHSA-2a, and the expression of transcription factor BmFTZ-F1, which belongs to 20E pathway and binds to CRE of BmFTZ-F1 , was inhibited by 20E. The results in this study demonstrate the relationship between hormones and BmCHSA-2a and provide the foundation for further study of possible target gene used for pest control.

     

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