文献类型：期刊文献

中文题名：白蜡虫泌蜡初期全长转录组测序及FAR在雌雄虫中蛋白表达分析

英文题名：Full-length Transcriptome Sequencing of Ericerus pela at the Early Stage of Wax Secretion and Protein Expression Analysis of FAR in Males and Females

作者：赵遵岭[1] 吴国星[2] 高熹[2] 于淑惠[3] 杨斌[4] 朱家颖[4] 杨璞[1]

第一作者：赵遵岭

机构：[1]中国林业科学研究院资源昆虫研究所,国家林业局资源昆虫培育与利用重点实验室,云南昆明650224;[2]云南农业大学植物保护学院,云南昆明650201;[3]昆明学院农学与生物技术学院,云南昆明650214;[4]西南林业大学生物多样性保护学院,云南省森林灾害预警与控制重点实验室,云南昆明650224

年份：2019

卷号：34

期号：5

起止页码：762-771

中文期刊名：云南农业大学学报：自然科学版

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；

基金：中央级公益性科研院所基本科研业务费专项(CAFYBB2017ZB005);国家自然科学基金项目(31572337);国家林业局林业公益性行业科研专项经费重大项目(201504302)

语种：中文

中文关键词：白蜡虫;白蜡合成基因;脂酰辅酶A还原酶;转录组测序;抗体制备;表达分析

外文关键词：Ericerus pela;genes responsible for wax synthesis;fatty acyl-CoA reductase;transcriptome sequencing;antibody preparation;expression analysis

分类号：Q78;S899.1

摘要：【目的】了解白蜡虫[Ericerus pela (Chavannes)]二龄雄虫泌蜡初期的基因表达,鉴定参与白蜡合成的脂酰辅酶A还原酶(fatty acyl-CoA reductase,far)基因,并明确FAR在白蜡虫雌雄虫中的蛋白表达情况。【方法】采用三代全长转录组测序,分析白蜡虫二龄雄虫泌蜡初期表达的基因,筛选获得与白蜡虫泌蜡相关的far基因;对关注的1个FAR制备单克隆抗体,通过Western-blot分析该FAR在白蜡虫雌雄虫中的表达情况。【结果】对白蜡虫泌蜡初期的二龄雄虫成功进行了转录组测序,共获得81个far基因,筛选获得与白蜡虫泌蜡一致的6个基因,本研究关注的far基因命名为far3;成功制备了FAR3的单克隆抗体,通过单克隆抗体分析了FAR3在白蜡虫雌雄虫中的表达情况,证实FAR3在二龄雄虫中大量表达,在雌虫中有微量表达。【结论】本研究通过对泌蜡初期的白蜡虫二龄雄虫进行三代全长转录组测序获得了与白蜡虫泌蜡相关的far基因;通过制备的FAR3单克隆抗体证实了FAR3在二龄雄虫分泌白蜡期高量表达,这为白蜡虫其他far基因功能研究以及探索白蜡虫泌蜡分子机理奠定了基础。
[Purpose]To understand the gene expression during the early stage of wax secretion in the second-instar males Ericerus pela, and to identify the fatty acyl-CoA reductase (far) gene involved in wax synthesis and the protein expression of FAR in females and males.[Method]Three generations of full-length transcriptome sequencing were used to analyze the genes expression of the second instar males of E. pela during the early stage of wax secretion, and the far genes related to wax secretion of E. pela were identified;monoclonal antibodies were prepared from a concerned FAR. The expression of the FAR in females and males was analyzed by using Western-blot.[Result]The third generations sequencing of full-length transcriptome was successfully performed on the second-instar males during the early stage of wax secretion of E. pela, a total of 81 far genes were obtained, and 6 far genes with expression profile consistent with the wax secretion of the insect were identified. One of the far genes was selected and named far3. The monoclonal antibody of FAR3 was successfully prepared. And the expression of FAR3 in males and females was analyzed by using the monoclonal antibody, it was confirmed that FAR3 was highly expressed in males during the second larval stage and slightly expressed in females.[Conclusion]In this study, the far genes related to wax secretion were obtained by three generations of full-length transcriptome sequencing of the second-instar males of E. pela during the early stage of wax secretion. The high expression of FAR3 in the second instar males during the wax secretion stage was confirmed by using the monoclonal antibodies of FAR3, which laid a foundation for the functional study of other far genes of E. pela and the exploration of the molecular mechanism of wax secretion of E. pela.