莲草直胸跳甲气味结合蛋白的基因克隆与功能初探

发布时间：2018-05-11 06:55

本文选题：莲草直胸跳甲 + 气味结合蛋白　；参考：《山西农业大学》2016年硕士论文

【摘要】：在长期的生物进化中,昆虫形成了灵敏的嗅觉系统,这对昆虫寻找食物、繁殖后代以及躲避天敌等都发挥着重要的作用。昆虫的嗅觉形成过程需要多种蛋白的参与,其中气味结合蛋白(Odorant binding proteins, OBPs)与外界气味的结合是昆虫嗅觉识别的第一步生化反应,起到初步筛选和鉴定气味分子的作用。莲草直胸跳甲(Agasicles hygrophila)是入侵杂草喜旱莲子草(Alternanthera philoxeroides)的专食性天敌昆虫。为揭示气味结合蛋白在莲草直胸跳甲寄主专一性中的作用,同时进一步了解气味结合蛋白在昆虫嗅觉系统中的功能作用,本试验开展了以下研究：1)利用Blastx t匕对从莲草直胸跳甲转录组中初步筛选鉴定出28个气味结合蛋白基因,可分为Classic OBPs、Minus-C OBPs 和 Atypical OBPs三类,分别为8、18和2个,大部分属于Minus-C OBPs。2)首次克隆了3个莲草直胸跳甲Classic OBPs (OBP12、OBP14 和 OBP24)基因,开放阅读框分别为351 bp、423 bp和408 bp,分别编码116、140和135个氨基酸,具有OBPs的典型特征,属于分子量较小的酸性可溶性蛋白质。3)通过荧光定量PCR方法,分析了三种气味结合蛋白的组织表达特征,结果表明三种气味结合蛋白均在雌雄虫的触角中表达量最高。OBP12和OBP14在雌虫触角中的表达水平高于雄虫,推测这两种蛋白可能与识别寄主植物挥发物和寻找产卵地点相关;而OBP24在雄虫触角中的表达水平高于雌虫,可能是识别雌虫散发的性信息素。另外,OBP12在足、头和翅中也有表达；OBP14除在触角表达,在头和足中均有表达;OBP24在头、足和翅中也有表达。4)构建了原核表达载体,成功诱导目的蛋白OBP12和OBP24的表达,为继续研究该蛋白的结构特点和生理功能奠定了良好的基础。
[Abstract]:In long-term biological evolution, insects form a sensitive olfactory system, which plays an important role in searching for food, breeding and avoiding natural enemies. The process of olfactory formation in insects requires the participation of many proteins. The binding of odour binding protein (Odorant binding proteins, OBPs) to external odors is the first biochemical reaction of olfactory recognition in insects and plays a role in screening and identifying odour molecules. Agasicles hygrophila is a special natural enemy insect of Alternanthera philoxeroides. In order to reveal the role of odor-binding protein in the host specificity of Lilidium chinensis, and to further understand the function of odor-binding protein in the olfactory system of insects. In this experiment, the following studies were carried out: (1) 28 odor-binding protein genes were preliminarily screened and identified by Blastx t from the transcriptional group of Liancao chinensis, which can be divided into three types: Classic OBPsMimus-C OBPs and Atypical OBPs, which are 80.18 and 2, respectively. Three OBP12OBP14 and OBP24) genes were cloned for the first time from Minus-C OBPs.2. the open reading frames were 351bp423bp and 408bp respectively, encoding 116140 and 135 amino acids, respectively, with the typical characteristics of OBPs. The tissue expression characteristics of three odor-binding proteins were analyzed by fluorescence quantitative PCR. The results showed that the expression level of OBP12 and OBP14 in the antennae of females and males were higher than that of males, which suggested that these proteins might be related to the identification of host plant volatiles and the search for spawning sites. The expression level of OBP24 in the antennae of males is higher than that of females, which may be a sex pheromone that recognizes the emission of females. In addition, OBP12 was expressed in foot, head and wings, except in antennae, OBP24 was expressed in head and foot, and OBP24 was expressed in head, foot and wings. A prokaryotic expression vector was constructed, which successfully induced the expression of OBP12 and OBP24. It lays a good foundation for the further study of the structural characteristics and physiological functions of the protein.
【学位授予单位】：山西农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S476;S451

【相似文献】