贵州医科大学学报

2018, v.43;No.208(01) 7-11

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兴义维蚋幼虫气管鳃特异表达候选基因筛选
Initial Screening of Candidate Genes Specifically Expressing in the Tracheal Gills of Simulium xingyiense Larvae

贾若;杨曜铭;寻慧;付小珍;曹梅;高维平;杨明;
JIA Ruo;YANG Yaoming;XUN Hui;FU Xiaozhen;CAO Mei;GAO Weiping;YANG Ming;Department of Biology,Guizhou Medical University;Key Laboratory of Medical Entomology,Guizhou Medical University;

摘要(Abstract):

目的:筛选兴义维蚋幼虫气管鳃发育早期特异表达的基因。方法:将发育成熟的1~7龄期的蚋幼虫气管鳃蛋白作为抗原免疫新西兰家兔,制备兴义维蚋气管鳃蛋白多克隆抗血清,用Western blot法分析气管鳃特异表达蛋白;以LC-MS/MS对气管鳃特异蛋白进行质谱分析,采用GO分析及手工检索Fly Base数据库分析蛋白组成。结果:发现63 k Da处存在整个幼虫期均表达的蛋白,对63~75 k Da蛋白进行质谱分析,发现具有几丁质结合特性的蛋白质Gasp和Cda4。结论:初步确定Gasp和Cda4基因可作为气管鳃早期发育特异表达候选基因。
Objective: To screen for the candidate genes specifically expressing in tracheal gills on early larval developmental stages of black fly Simulium xingyiense. Methods: The polyclonal antiserum of tracheal gills of S. xingyiense was prepared. Specific protein expression of tracheal gills during larval stages was comparatively analyzed using Western Blot. Mass spectrometry of tracheal gill-specific protein complex was analyzed by LC-MS/MS. Functions of derived proteins were annotated with aid of GO annotation and Fly Base. Results: It showed that there existed proteins with molecular weight about 63 k Da expressed during whole larval stages. Two proteins,Gasp and chitin deacetylase-like 4( Cda4),with molecular weight between 63 k Da and 75 k Da,were identified as chitin binding proteins by mass spectrometry. Conclusions: It is reasonable to select Gasp and Cda4 genes as candidates expressing specifically in the early developmental stages of tracheal gills of S. xingyiense larvae.

关键词(KeyWords): 兴义维蚋;气管鳃;多克隆抗血清;质谱鉴定;GO分析
Simulium xingyiense;tracheal gill;polyclonal antiserum;mass spectrometry;GO analysis

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基金项目(Foundation): 国家自然科学基金项目(No30660021);; 贵州省社会发展攻关项目[黔科合SY(2012)3083];; 贵州医科大学科技基金(校重点实验室2016003);; 贵州省科技计划[黔科合LG(2012)022]

作者(Author): 贾若;杨曜铭;寻慧;付小珍;曹梅;高维平;杨明;
JIA Ruo;YANG Yaoming;XUN Hui;FU Xiaozhen;CAO Mei;GAO Weiping;YANG Ming;Department of Biology,Guizhou Medical University;Key Laboratory of Medical Entomology,Guizhou Medical University;

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