本文選題：轉(zhuǎn)錄組 + 第二代高通量測序��； 參考：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：嗅覺感受系統(tǒng)在昆蟲取食、尋找寄主、躲避天敵、交配和產(chǎn)卵等一系列生命活動中發(fā)揮了重要作用；在昆蟲嗅覺感受系統(tǒng)中,氣味分子結(jié)合蛋白(odorant-binding protein, OBP)扮演著重要的角色。本研究利用第二代高通量測序技術(shù)在亞洲小車蝗(Oedaleus asiaticus)觸角中首次鑒定出15個編碼OBP的基因,命名為OBP1-15,并對其表達譜進行了測定與分析,此外,又對亞洲小車蝗OBP與包括東亞飛蝗在內(nèi)的其它六種昆蟲的OBP進行了系統(tǒng)發(fā)育分析。主要結(jié)果如下：1.亞洲小車蝗雌、雄成蟲觸角轉(zhuǎn)錄組測序、數(shù)據(jù)分析及氣味分子結(jié)合蛋白基因的鑒定。通過對亞洲小車蝗成蟲雌、雄觸角分別進行轉(zhuǎn)錄組測序,獲得總clean reads數(shù)分別為137,426,706和127,822,838;總clean堿基數(shù)分別為12,368,403,540 nt和11,504,055,420 nt,均超過10GB,經(jīng)過軟件組裝后,共獲得61,629條all-unigene,其中有26,064(42.30%)條all-unigene獲得了功能注釋信息。首次在亞洲小車蝗成蟲觸角中鑒定出15個OBP基因,分析發(fā)現(xiàn)它們均具有全長開放閱讀編碼框(open reading frame, ORF)。2.亞洲小車蝗氣味分子結(jié)合蛋白基因表達譜測定。通過實時熒光定量PCR (quantitative real-time PCR, qRT-PCR)實驗,發(fā)現(xiàn)只有OBP7在雌觸角中的表達量要遠大于在雄觸角中的表達量；有9個OBP在雄觸角中的表達量要大于在雌觸角中的表達量。通過半定量PCR實驗,發(fā)現(xiàn)OBP9和OBP10只特異性的表達于觸角,而其它OBP則在觸角、口器或翅膀等化學(xué)感受組織中表達。3.亞洲小車蝗OBP與其他昆蟲OBP系統(tǒng)發(fā)育分析。通過對亞洲小車蝗15個OBP與其它來自于六個不同目的六種昆蟲OBP氨基酸序列之間構(gòu)建的進化樹進行系統(tǒng)發(fā)育分析,發(fā)現(xiàn)亞洲小車蝗的大部分OBP都與東亞飛蝗(Locusta migratoria) OBP親緣關(guān)系最近,但是亞洲小車蝗有些OBP并不是與東亞飛蝗親緣關(guān)系最近。這表明二者的OBP基因最初由同一個祖先基因進化而來,但是為了適應(yīng)不同的生態(tài)環(huán)境,各自進化出了獨特的化學(xué)感受系統(tǒng)。
[Abstract]:Olfactory sensing system plays an important role in a series of life activities, such as feeding, finding hosts, avoiding natural enemies, mating and spawning, etc. In insect olfactory sensing system, odor molecule binding protein odorant-binding protein (OBPP) plays an important role. In this study, the second generation high-throughput sequencing technique was used to identify for the first time 15 genes encoding OBP in the antennae of Oedaleus asiaticus, named OBP1-15, and their expression profiles were determined and analyzed. The phylogenetic analysis of OBP and six other insects, including migratory locust in East Asia, was also carried out. The main results are as follows: 1. Transcriptome sequencing, data analysis and identification of odour molecular binding protein gene in female and male adults. By sequencing the female and male antennae, the total clean reads numbers were 137426706 and 127822838, and the total clean base numbers were 12368403540 NT and 11504055420 NT, respectively. A total of 61629 all-unigenetics were obtained, of which 26064 and 42.30) all-unigene obtained functional annotation information. For the first time, 15 OBP genes were identified from the antennae of Acridostoma asiatica. It was found that all of them had full-length open reading frame, ORF.2. Detection of molecular binding protein gene expression profile of Asiatic locust. By real-time quantitative PCR quantitative real-time PCR (qRT-PCR) experiment, it was found that only OBP7 was expressed in the female antennae and 9 OBP in the male antennae were higher than those in the female antennae. By semi-quantitative PCR assay, it was found that OBP9 and OBP10 expressed only in the antennae, while other OBP expressed .3in chemosensory tissues such as antennae, mouthparts or wings. Phylogenetic analysis of OBP and other insects OBP. Phylogenetic analysis of the phylogenetic tree constructed between 15 OBP and other OBP amino acid sequences of six insect species from six different purposes was carried out. It was found that most of the OBP was closely related to Locusta migratoria OBP, but some OBP were not closely related to the Asian migratory locust. This indicates that the OBP gene of the two genes was originally evolved from the same ancestor gene, but in order to adapt to different ecological environments, they have evolved a unique chemical sensing system.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S433.2;Q78

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