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  本文選題：蓮草直胸跳甲 + 氣味結(jié)合蛋白��； 參考：《山西農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：在長期的生物進(jìn)化中,昆蟲形成了靈敏的嗅覺系統(tǒng),這對昆蟲尋找食物、繁殖后代以及躲避天敵等都發(fā)揮著重要的作用。昆蟲的嗅覺形成過程需要多種蛋白的參與,其中氣味結(jié)合蛋白(Odorant binding proteins, OBPs)與外界氣味的結(jié)合是昆蟲嗅覺識別的第一步生化反應(yīng),起到初步篩選和鑒定氣味分子的作用。蓮草直胸跳甲(Agasicles hygrophila)是入侵雜草喜旱蓮子草(Alternanthera philoxeroides)的專食性天敵昆蟲。為揭示氣味結(jié)合蛋白在蓮草直胸跳甲寄主專一性中的作用,同時進(jìn)一步了解氣味結(jié)合蛋白在昆蟲嗅覺系統(tǒng)中的功能作用,本試驗開展了以下研究：1)利用Blastx t匕對從蓮草直胸跳甲轉(zhuǎn)錄組中初步篩選鑒定出28個氣味結(jié)合蛋白基因,可分為Classic OBPs、Minus-C OBPs 和 Atypical OBPs三類,分別為8、18和2個,大部分屬于Minus-C OBPs。2)首次克隆了3個蓮草直胸跳甲Classic OBPs (OBP12、OBP14 和 OBP24)基因,開放閱讀框分別為351 bp、423 bp和408 bp,分別編碼116、140和135個氨基酸,具有OBPs的典型特征,屬于分子量較小的酸性可溶性蛋白質(zhì)。3)通過熒光定量PCR方法,分析了三種氣味結(jié)合蛋白的組織表達(dá)特征,結(jié)果表明三種氣味結(jié)合蛋白均在雌雄蟲的觸角中表達(dá)量最高。OBP12和OBP14在雌蟲觸角中的表達(dá)水平高于雄蟲,推測這兩種蛋白可能與識別寄主植物揮發(fā)物和尋找產(chǎn)卵地點相關(guān);而OBP24在雄蟲觸角中的表達(dá)水平高于雌蟲,可能是識別雌蟲散發(fā)的性信息素。另外,OBP12在足、頭和翅中也有表達(dá)；OBP14除在觸角表達(dá),在頭和足中均有表達(dá);OBP24在頭、足和翅中也有表達(dá)。4)構(gòu)建了原核表達(dá)載體,成功誘導(dǎo)目的蛋白OBP12和OBP24的表達(dá),為繼續(xù)研究該蛋白的結(jié)構(gòu)特點和生理功能奠定了良好的基礎(chǔ)。
[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.
【學(xué)位授予單位】：山西農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S476;S451

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