Mmu-143调控牙鲆Tyrp1a基因表达及牙鲆白化皮肤细胞系的建立

发布时间：2018-08-24 20:00

【摘要】：牙鲆是我国北方重要的海水养殖鱼类,苗种生产中普遍存在体色白化现象,严重影响了苗种品质和市场价值。为了开展牙鲆白化分子调控机制方面的研究,本论文以牙鲆白化和原色样品为材料,在获得转录组及microRNA(miRNA)深度测序数据的基础上,拟从转录和转录后水平上研究牙鲆白化发生机制。通过mRNA与miRNA联合分析,筛选并鉴定牙鲆白化关键miRNA mmu-143及其靶基因Tyrp1。通过对牙鲆白化靶基因和miRNA mmu-143的表达模式验证分析,进行miRNA mmu-143在牙鲆白化发生过程中调控机制的研究;本实验还用甲状腺激素T3处理牙鲆仔鱼,结果导致牙鲆白化比例增加,且上调表达基因Complement component C8beta chain(c8b)、Complement component C9(c9)、Myoblast determination protein1 homolog(myod)和下调表达基因ATP-dependent 6-phosphofructokinase(pfkm)、Melanotransferrin(mfi2)、Somatotropin(gh)的定量结果与高通量测序结果一致;以牙鲆原色和白化皮肤为材料,分别建立皮肤细胞系并命名为JFSK_wt和JFSK_alb,并进行了细胞生长曲线、核型分析、病毒敏感性、转染和白化相关基因表达情况研究这一系列实验,结果表明这两株细胞系可以为目标基因或microRNA的功能研究提供有效的细胞平台。本论文研究结果为深入揭示牙鲆白化发生的分子机制提供了重要的基础资料。
[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).
【学位授予单位】：上海海洋大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S917.4

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