斑马鱼nrf基因时空表达分析及其在调控胰外分泌酶原基因表达中的作用研究

发布时间：2019-05-29 03:40

【摘要】：通过GenBank搜索发现,斑马鱼中有6个nrf同源基因,即nfe2、nrf1a、nrf1b、nrf2a、nrf2b和nrf3。为了研究6个nrf在物种之间的关系,实验对6个Nrf蛋白进行系统发育树分析,结果显示6个Nrf蛋白的分子进化趋势与物种进化趋势一致,尽管在硬骨鱼类中由于基因组倍增的原因,nrf1和nrf2基因各产生了两个拷贝,但Nrf蛋白仍然是比较保守的。此外,实验选取斑马鱼胚胎发育十个不同时期(1 cell、2 cell、3.5 hpf、6 hpf、12 hpf、24 hpf、36 hpf、48 hpf、72 hpf、96 hpf),对其表达模式进行了详细的研究,结果显示:6个nrf在一细胞期都有转录信号,且胚胎发育早期(48 hpf之前)全身广泛表达。72 hpf后,nrf1b、nrf2a、nrf2b、nrf3和nfe2基因的表达部位主要集中在头部、部分躯干、肠处,nrf2b还在脊髓处有微弱的表达。总体来说,6个nrf基因的表达部位基本重叠。研究还利用双荧光素酶检测系统检测了6个Nrf蛋白对胰外分泌酶原基因(tryl)转录的调控,结果显示nrf1a、nrf2a、nrf2b与nfe2的过表达能使tryl的表达上调,而nrf1b与nrf3的过表达能抑制tryl的表达。研究结果将为深入研究斑马鱼6个nrf基因的功能叠加、互补及其功能歧化奠定基础。
[Abstract]:Through GenBank search, it was found that there were six nrf homologous genes in zebrafish, namely nfe2,nrf1a,nrf1b,nrf2a,nrf2b and nrf3.. In order to study the relationship of six nrf among species, the phylogenetic tree analysis of 6 Nrf proteins was carried out. The results showed that the molecular evolution trend of 6 Nrf proteins was consistent with that of species. Although two copies of nrf1 and nrf2 genes were produced in osseous fish due to genome doubling, Nrf protein was still conserved. In addition, ten different stages of embryonic development of zebrafish (1 cell,2 cell,3.5 hpf,6 hpf,12 hpf,24 hpf,36 hpf,48 hpf,72 hpf,96 hpf),) were selected to study the expression pattern of zebrafish embryo in detail. The results showed that all 6 nrf had transcriptional signals in one cell stage, and they were widely expressed in the whole body at the early stage of embryonic development (before 48 hpf). After 72 hpf, the expression of nrf1b,nrf2a,nrf2b,nrf3 and nfe2 genes was mainly concentrated in the head, part of the torso and intestine. Nrf2b was also weakly expressed in the spinal cord. On the whole, the expression sites of six nrf genes basically overlapped. The regulation of pancreatic exocrine proto gene (tryl) transcription by six Nrf proteins was also detected by double luciferase detection system. The results showed that the overexpression of nrf1a,nrf2a,nrf2b and nfe2 could up-regulate the expression of tryl. The overexpression of nrf1b and nrf3 could inhibit the expression of tryl. The results will lay a foundation for further study on the functional superposition, complementarity and functional ambiguity of six nrf genes in zebrafish.
【作者单位】：华中师范大学生命科学学院湖北省遗传调控与整合生物学重点实验室;国家海洋局海洋生物遗传资源重点实验室;
【基金】：国家自然科学基金项目(31071997,31272645) 国家海洋局海洋生物遗传资源重点实验室开放课题(HY201606)资助~~
【分类号】：Q78

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