本文選題：日本鰻鱺 + Dmrt2基因　； 參考：《上海海洋大學(xué)》2017年碩士論文

【摘要】：Dmrt(double-sex and mab3-relatated transcription factor)基因是與性別決定與性腺發(fā)育有關(guān)的一個(gè)基因家族,同時(shí)對(duì)某些器官的發(fā)育和功能維持也起著重要作用。隨著分子手段的進(jìn)步,Dmrt基因家族的研究也在日益增進(jìn),最新的研究表明Dmrt2基因主要參與胚胎的發(fā)育和配子的形成。本實(shí)驗(yàn)結(jié)果表明日本鰻鱺Dmrt2基因表達(dá)蛋白僅含有一個(gè)功能結(jié)構(gòu)域,即DM結(jié)構(gòu)域。其Dmrt2蛋白的分子量為46.82kD,分子式為C2037H3281N595O617S26,總原子數(shù)為6556, Dmrt2蛋白分子的氨基酸組成主要包括以下幾種:Ser (S),11.1%; Leu (L),10.8%; Lys (K) 7.5%和Trp (W),0.7%。利用模型軟件預(yù)測(cè)日本鰻鱺Dmrt2基因表達(dá)蛋白的二級(jí)結(jié)構(gòu)包含14個(gè)α-螺旋,17個(gè)β-折疊并具有多個(gè)卷曲。通過軟件比較不同物種的Dmrt2基因表達(dá)蛋白的結(jié)構(gòu)域,發(fā)現(xiàn)其相對(duì)保守。通過Race技術(shù)對(duì)Dmrt2基因的5'和3'序列進(jìn)行擴(kuò)增并拼接獲得Dmrt2基因的cDNA全長(zhǎng)為1499bp。通過對(duì)氨基酸序列的分析和同源性對(duì)比顯示,日本鰻鱺和斑馬魚(Danio rerio)及尼羅羅非魚(Oreochromis niloticus)之間具有高同源性。為了研究Dmrt2在日本鰻鱺的性別分化中的功能以及在個(gè)體發(fā)育中的作用,采用RT-PCR方法對(duì)Dmrt2基因在不同組織中的表達(dá)進(jìn)行了分析。采用FISH分析驗(yàn)證Dmrt2在性腺中的表達(dá)情況。本研究結(jié)果顯示,Dmrt2基因在多種組織中表達(dá),并且在卵巢中相對(duì)較高,在精巢,肌肉和肝組織中弱表達(dá)。對(duì)于用E2 (雌二醇-17β)處理的日本鰻鱺,Dmrt2基因在卵巢和肝臟中表達(dá)較高,而在腦和肌肉組織中弱表達(dá)。熒光原位雜交結(jié)果表明Dmrt2基因的雜交信號(hào)只在卵巢中的卵泡細(xì)胞中可見,在性成熟的精巢中沒有發(fā)現(xiàn)雜交信號(hào)。這些數(shù)據(jù)將為日本鰻鱺性別分化的進(jìn)一步提供理論基礎(chǔ)。
[Abstract]:The double-sex and mab3-relatated transcription factor gene is a family of genes related to sex determination and gonadal development. It also plays an important role in the development and functional maintenance of some organs. With the development of molecular techniques, the study of Dmrt gene family is increasing. The latest research shows that Dmrt2 gene is mainly involved in embryo development and gamete formation. The results showed that the Dmrt2 gene expression protein of Japanese eel contained only one functional domain, DM domain. The molecular weight of Dmrt2 protein is 46.82kD. the molecular formula is C2037H3281N595O617S26.The total atomic number of Dmrt2 protein is 6556. The amino acid composition of Dmrt2 protein mainly consists of the following kinds of: Ser Schion 11. 1; Leu L10.8; Lys K1) 7.5%; The secondary structure of Dmrt2 gene expressed protein in Japanese eel Anguilla japonica contained 14 偽 -helix, 17 尾 -folds and multiple curls. The domain of Dmrt2 gene expressed in different species was found to be relatively conserved by software. The 5 'and 3' sequences of Dmrt2 gene were amplified by Race technique and the full-length cDNA of Dmrt2 gene was obtained to be 1499bp. Amino acid sequence analysis and homology comparison showed that there was a high homology between Japanese eel and Danio Rerioo) and Oreochromis niloticus of Oreochromis niloticus. In order to study the function of Dmrt2 in sex differentiation and ontogeny of Japanese eel, the expression of Dmrt2 gene in different tissues was analyzed by RT-PCR. Fish analysis was used to verify the expression of Dmrt2 in gonad. The results showed that the Dmrt2 gene was expressed in a variety of tissues and was relatively high in the ovary and weakly expressed in the testis, muscles and liver. The expression of Dmrt2 gene in Japanese eel (Anguilla japonica) treated with E2 (estradiol-17 尾) was higher in ovary and liver, but weak in brain and muscle. The results of fluorescence in situ hybridization showed that the hybridization signal of Dmrt2 gene was only found in follicular cells of ovary, but not in the testis of sexual maturity. These data will provide theoretical basis for sex differentiation of Japanese eel.
【學(xué)位授予單位】：上海海洋大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S917.4

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本文編號(hào)：2023655