本文選題：中華蜜蜂　切入點：氣味受體基因　出處：《山西農(nóng)業(yè)大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：本研究以中華蜜蜂為研究對象,對其三個傳統(tǒng)氣味受體基因AcerOr57、AcerOrl39和AcerOrl67進(jìn)行克隆、序列分析和定量表達(dá)分析,研究結(jié)果如下：1利用RT-PCR法獲得了中華蜜蜂氣味受體基因AcerOr57、AcerOrl39和AcerOr167的cDNA序列(GenBank登錄號分別為：KT716393、KT239370和KT239369),包括完整的開放閱讀框,分別編碼407、387和436個氨基酸。生物信息學(xué)表明,這3個氣味受體均為疏水性蛋白;分別具有6個、7個和5個跨膜結(jié)構(gòu)域；均無信號肽和糖基化位點;AcerOr57和AcerOr139的結(jié)構(gòu)不穩(wěn)定,AcerOr167的結(jié)構(gòu)較穩(wěn)定;這與昆蟲傳統(tǒng)氣味受體的結(jié)構(gòu)特征相符合。2通過在NCBI數(shù)據(jù)庫中Blastp搜索,分別找到與AcerOr57、AcerOrl39和AcerOr167的同源的序列,多序列比對發(fā)現(xiàn)AcerOr57與西方蜜蜂AmelOr57的相似性最高,為97%,且與AmelOr55、AmelOr56和AmelOr58均具有較高的相似性,推測AcerOr55、AcerOr56、AcerOr57和AcerOr58可能是蜜蜂在進(jìn)化過程中通過基因復(fù)制產(chǎn)生的同源基因,且有可能串聯(lián)排列在同一染色體上;AcerOr139也與西方蜜蜂的多個氣味受體具有較高的相似性,其中與西方蜜蜂的假基因AmelOr139的相似性最高,為73%,對于AcerOr139是否也喪失原有功能,產(chǎn)生新的功能需要深入的研究；AcerOr167與膜翅目昆蟲氣味受體具有較高的的相似性,與西方蜜蜂AmelOr167的相似性最高,為94%,說明它們是直系同源基因。3利用實時熒光定量技術(shù)對AcerOr57, AcerOrl39和AcerOrl67在工蜂羽化后不同組織(觸角、頭、胸、腹、足、翅)和不同發(fā)育階段(1、5、10、15、20、25和30日齡)中的相對表達(dá)量進(jìn)行檢測。結(jié)果表明,AcerOr57和AcerOr167 mRNA在工蜂各組織中均有表達(dá),其中,在觸角中的表達(dá)量極顯著高于其他組織(P0.01); AcerOr139 mRNA也在工蜂各組織中均表達(dá),但在不同的發(fā)育階段表達(dá)特性不同,在1、5、25和30日齡時觸角的表達(dá)量最高,10、15和20日齡時頭部的表達(dá)量最高,推測這3個氣味受體與蜜蜂的嗅覺識別作用有關(guān)。AcerOr57 mRNA在工蜂羽化后1日齡時表達(dá)量最高,10日齡最低,之后呈逐漸上升的趨勢,推測其可能與中蜂出巢采集,識別花香氣味有關(guān);AcerOr139 mRNA在工蜂羽化后5、25和30日齡時均出現(xiàn)相對較高的表達(dá),AcerOr167 mRNA在工蜂羽化后5日齡和20日齡時呈現(xiàn)相對較高的表達(dá),推測這兩個基因在中蜂識別花粉和花蜜氣味分子過程中起著重要的作用。另外,AcerOr167 mRNA在工蜂整個發(fā)育階段的表達(dá)量明顯高于AcerOr57和AcerOr139,推測AcerOr167可能在中蜂的嗅覺系統(tǒng)中發(fā)揮著重要的作用
[Abstract]:In this study, the bees as the research object, the three traditional odorant receptor gene AcerOr57, AcerOrl39 and AcerOrl67 were cloned, expressed sequence analysis and quantitative analysis, the research results are as follows: 1 by using the RT-PCR method to obtain the bee odorant receptor gene AcerOr57, AcerOrl39 and AcerOr167 cDNA sequence (GenBank accession number: KT716393, KT239370 and KT239369), including the complete open reading frame encoding 407387 and 436 amino acids respectively. Bioinformatics showed that these 3 odorant receptors were hydrophobic protein; respectively with 6, 7 and 5 transmembrane domains; no signal peptide and glycosylation sites; AcerOr57 and AcerOr139 the instability of the structure of AcerOr167 is stable; and the traditional insect odorant receptor structure with.2 in the NCBI database by Blastp search, were found with AcerOr57, AcerOrl39 and AcerOr167 The homologous sequence, sequence comparison showed that the highest similarity, AcerOr57 and Western bees AmelOr57 was 97%, and the AmelOr55, AmelOr56 and AmelOr58 were similar, higher AcerOr55, AcerOr56, AcerOr57 and AcerOr58 may be the bees in the process of evolution by gene duplication homologous genes, and there may be arranged in series on the same chromosome; a plurality of odorant receptor AcerOr139 and Western bees have higher similarity, the highest similarity of pseudogene AmelOr139 with Western bees was 73%, for AcerOr139 is the loss of the original function, which further research need a new similarity function; AcerOr167 and odorant receptors with Hymenoptera the higher, and the highest, similar to Western bees AmelOr167 was 94%, indicating that they are orthologs of.3 using real-time fluorescence quantitative AcerOr57 technology of AcerOrl. 39 and AcerOrl67 in different tissues after emergence (worker antennae, head, chest, abdomen, feet, wings) and different developmental stages (1,5,10,15,20,25 and 30 days of age) relative expression quantity in the test. The results showed that AcerOr57 and AcerOr167 mRNA were expressed in various tissues in the expression of the worker, the antenna. Was significantly higher than that of other organizations (P0.01); AcerOr139 mRNA was also expressed in tissues of workers, but the expression in different developmental stages of different characteristics, expression of antenna in 1,5,25 and at the age of 30, the highest 10,15, and at the age of 20, the head of the highest expression, speculated that the 3 odor receptors and bees the role of.AcerOr57 in olfactory mRNA workers after emergence at the age of 1, the highest expression level at 10 days of age, the lowest, after gradually rising trend, suggesting that it may be with the bee nest collection, identification of floral scent; AcerOr139 mRNA and 30 5,25 after emergence in worker Relatively high expression showed age, AcerOr167 mRNA in worker 5 after emergence and 20 days of age showed a relatively higher level, it is suggested that the two gene plays an important role in the identification of bee pollen and nectar odor molecules in the process. In addition, the AcerOr167 expression of mRNA in the whole growth stage of worker the amount was significantly higher than that of AcerOr57 and AcerOr139, AcerOr167 speculated that the olfactory system may in the bee play an important role in

【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】：S891;Q78

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