Trypsin is one of the most diverse and widely studied protease hydrolases. However, the diversity and characteristics of the Trypsin superfamily of genes have not been well understood, and their role in insecticide resistance is yet to be investigated. In this study, a total of 342 Trypsin genes were identified and classified into seven families based on homology, characteristic domains and phylogenetics in , and the LY-Domain and CLECT-Domain families are specific to the species. Four Trypsin genes, (, , , ) were identified to be associated with pyrethroid resistance based on transcriptome analyses of three field resistant populations and qRT-PCR validation, and the knock-down of these genes significantly decrease the pyrethroid resistance of based on RNAi. The activity of Astry43a can be reduced by five selected insecticides (indoxacarb, DDT, temephos, imidacloprid and deltamethrin); and however, the Astry43a could not directly metabolize these five insecticides, like the trypsin NYD-Tr did in earlier reports. This study provides the overall information frame of Trypsin genes, and proposes the role of Trypsin genes to insecticide resistance. Further researches are necessary to investigate the metabolism function of these trypsins to insecticides.

中文翻译：

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基于组学和功能分析的疟疾媒介中华按蚊胰蛋白酶基因特征及其在杀虫剂抗药性中的作用

胰蛋白酶是最多样化且研究最广泛的蛋白酶水解酶之一。然而，胰蛋白酶超家族基因的多样性和特征尚未得到充分了解，其在杀虫剂抗性中的作用还有待研究。本研究共鉴定了342个胰蛋白酶基因，根据同源性、特征域和系统发育学将其分为7个家族，其中LY-Domain和CLECT-Domain家族是物种特有的。基于三个田间抗性群体的转录组分析和 qRT-PCR 验证，确定四个胰蛋白酶基因 (, , , ) 与拟除虫菊酯抗性相关，并且这些基因的敲低显着降低了基于 RNAi 的拟除虫菊酯抗性。 Astry43a的活性可被五种选定的杀虫剂（茚虫威、滴滴涕、替美磷、吡虫啉和溴氰菊酯）降低；然而，Astry43a 无法像早期报告中的胰蛋白酶 NYD-Tr 那样直接代谢这五种杀虫剂。本研究提供了胰蛋白酶基因的整体信息框架，并提出了胰蛋白酶基因在杀虫剂抗性中的作用。需要进一步的研究来调查这些胰蛋白酶对杀虫剂的代谢功能。