斑马鱼Brd3b调控消化器官发育的分子机制研究

项目名称： 斑马鱼Brd3b调控消化器官发育的分子机制研究

项目编号： No.31271557

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 黄红辉

作者单位： 西南大学

项目金额： 88万元

中文摘要： 消化器官是人体内的重要器官，研究消化器官发育对于进一步认识临床疾病、创新治疗方法有着重要意义。保守的分子机制和多样化的遗传手段使得斑马鱼成为理想的研究消化器官发育的模式动物。Brd3可特异性识别乙酰化组蛋白而调控基因表达，brd3b是我们鉴定的一个富集于斑马鱼胚胎消化器官内表达的基因，它在消化器官发育中的功能未见报道。敲降此基因的表达导致斑马鱼胚胎消化器官的发育严重缺陷，蛋白共沉淀的结果显示Brd3b与先前报道的另一消化器官富集因子核仁蛋白Def 有交互作用。本课题拟通过以下实验研究brd3b基因在消化器官发育中的功能与分子机制：1）TALEN法建立brd3b突变体，2）brd3b基因缺陷导致的消化器官发育缺陷的表型分析，3）Brd3b与Def的关系研究，4）Brd3b抗体染色质免疫共沉淀候选基因的鉴定，5）Brd3b调控的靶基因的鉴定，6）Brd3b靶基因在消化器官发育中的功能研究。

中文关键词： 斑马鱼；消化器官；Brd3b；Ltv1；核糖体病

英文摘要： Digestive organs play essential roles in human body, and studies of development of digestive organs implicate clinic applications. The conserved molecular mechanisms and powerful genetics of zebrafish make it as an ideal animal model to study digestive organogenesis.Brd3b, a member of BET (bromodomains and extraterminal) family, has two functionally conserved bromodomains, which can recognize and bind to acetyl lysine residues in histones to regulate downstream gene expression. brd3b was identified in our lab as a digestive organ enriched factor in an EST expression pattern screen. Morphants wiped out brd3b have severe defects in digestive organogenesis. Brd3b was also found to interact with Def, another digestive organs enriched nuclear factor. To elucidate the molecular mechanism of Brd3b in digestive organogenesis, we propose to perform studies in two ways: developmental study and molecular mechanism study. To obtain brd3b mutant line, the TALEN method will be used, and phenotype characterization will be performed in mutant lines. To identify target genes of Brd3b, ChIP-PCR will be used to test the candidate genes from Brd3b ChIP-Seq and RNA-Seq of mutant via siblings. Of course, those Brd3b target genes will be functional analyzed by gene knock-down or mRNA overexpression in brd3b mutants. We also want to fi

英文关键词： zebrafish；digestive organ；Brd3b；Ltv1；ribosomopathies