【摘要】：牙鲆是我國(guó)北方重要的海水養(yǎng)殖魚類,苗種生產(chǎn)中普遍存在體色白化現(xiàn)象,嚴(yán)重影響了苗種品質(zhì)和市場(chǎng)價(jià)值。為了開展牙鲆白化分子調(diào)控機(jī)制方面的研究,本論文以牙鲆白化和原色樣品為材料,在獲得轉(zhuǎn)錄組及microRNA(miRNA)深度測(cè)序數(shù)據(jù)的基礎(chǔ)上,擬從轉(zhuǎn)錄和轉(zhuǎn)錄后水平上研究牙鲆白化發(fā)生機(jī)制。通過mRNA與miRNA聯(lián)合分析,篩選并鑒定牙鲆白化關(guān)鍵miRNA mmu-143及其靶基因Tyrp1。通過對(duì)牙鲆白化靶基因和miRNA mmu-143的表達(dá)模式驗(yàn)證分析,進(jìn)行miRNA mmu-143在牙鲆白化發(fā)生過程中調(diào)控機(jī)制的研究;本實(shí)驗(yàn)還用甲狀腺激素T3處理牙鲆仔魚,結(jié)果導(dǎo)致牙鲆白化比例增加,且上調(diào)表達(dá)基因Complement component C8beta chain(c8b)、Complement component C9(c9)、Myoblast determination protein1 homolog(myod)和下調(diào)表達(dá)基因ATP-dependent 6-phosphofructokinase(pfkm)、Melanotransferrin(mfi2)、Somatotropin(gh)的定量結(jié)果與高通量測(cè)序結(jié)果一致;以牙鲆原色和白化皮膚為材料,分別建立皮膚細(xì)胞系并命名為JFSK_wt和JFSK_alb,并進(jìn)行了細(xì)胞生長(zhǎng)曲線、核型分析、病毒敏感性、轉(zhuǎn)染和白化相關(guān)基因表達(dá)情況研究這一系列實(shí)驗(yàn),結(jié)果表明這兩株細(xì)胞系可以為目標(biāo)基因或microRNA的功能研究提供有效的細(xì)胞平臺(tái)。本論文研究結(jié)果為深入揭示牙鲆白化發(fā)生的分子機(jī)制提供了重要的基礎(chǔ)資料。
[Abstract]:Paralichthys olivaceus is an important mariculture fish in the north of China. In order to study the molecular regulation mechanism of albinism in Paralichthys olivaceus, the albino and primary color samples of Paralichthys olivaceus were used as materials, and the transcriptome and microRNA (miRNA) depth sequencing data were obtained. To study the mechanism of albinism in Paralichthys olivaceus at the transcriptional and post-transcriptional levels. By combined analysis of mRNA and miRNA, the key miRNA mmu-143 and its target gene Tyrp1. of albino Paralichthys olivaceus were screened and identified. The regulation mechanism of miRNA mmu-143 during albinism was studied by verifying and analyzing the expression pattern of albino target gene and miRNA mmu-143 in Paralichthys olivaceus, and the ratio of albinism of flounder Paralichthys olivaceus was increased after treated with thyroid hormone T3. The quantitative results of up-regulated Complement component C8beta chain (c8b) complement component C9 (c9) Myoblast determination protein1 homolog (myod) and down-regulated ATP-dependent 6-phosphofructokinase (pfkm) ATP-dependent 6-phosphofructokinase (pfkm) Melanotransferrin (mfi2) Somatotropin (gh) were consistent with those of high throughput sequencing, and the primary color and albino skin of Paralichthys olivaceus were used as materials. Skin cell lines were established and named JFSK_wt and JFSK_alb, and cell growth curve, karyotype analysis, virus sensitivity, transfection and albinism related gene expression were studied. The results showed that the two cell lines could provide an effective cell platform for the functional study of target gene or microRNA. The results of this paper provide important basic data for revealing the molecular mechanism of albinism in Paralichthys olivaceus (Paralichthys olivaceus).
【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)：2201877