Jhdm1d论文小鼠新基因Jhdm1d的印记鉴定和表达谱分析.doc

Jhdm1d论文：小鼠新基因Jhdm1d的印记鉴定和表达谱分析 【中文摘要】Jhdm1d命名为组蛋白去甲基化酶1同系物D,定位于小鼠6号染色体,编码由940个氨基酸构成的组蛋白去甲基化酶,属于包含JmjC结构域的Jumonji蛋白家族。JmjC结构域被认为可能是一类新的组蛋白去甲基化酶共同基序,对H3K9me2和H3K27me2有去甲基化作用,研究发现Jumonji家族中的有些蛋白参与转录抑制和染色质调节过程。为了探索Jhdm1d基因在小鼠胚胎不同发育阶段的表达与组织发育的关系,本课题对Jhdm1d基因在mRNA水平上的时空表达谱进行了初步分析,为进一步探索该基因编码的组蛋白去甲基化酶在小鼠发育过程中的功能奠定理论基础。本课题选取小鼠胚胎为实验材料,利用原位杂交技术对Jhdm1d基因进行表达谱分析。全胚胎原位杂交数据表明该基因在E (embryonic day) 9.5、E10.5和E11.5天的小鼠全胚胎脑组织中持续高表达,此???,在E9.5天胚胎的心脏和E11.5天胚胎的体节中都有较强的信号表达;胚胎组织切片的原位杂交数据表明,Jhdm1d基因在E12.5天小鼠的脑、肝脏、心脏、脊椎软骨原基、嗅上皮、舌等器官和组织中均有不同水平的表达,其中脑、脊椎软骨原基、肝中表达水平较高;在E15.5天胚胎组织切片中,该基因在脑、肝、脊柱软骨原基、肾、肾上腺、肺、嗅上皮、舌、肋软骨、心等器官和组织中均有不同水平的表达,其中肝脏组织里表达信号最强,脑次之,脊柱软骨原基、肋软骨和肾上腺中也有较强表达。此外,选用E15.5天BFF1 (C57B L/6J♀×FVB♂)脑组织DNA和cDNA为材料,利用SNP (Single Nucleotide Polymorphism)位点直接测序法初步对Jhdm1d基因进行印记分析,结果表明Jhdm1d在E15.5天BFF1小鼠脑组织中为非印记基因。研究结果表明Jhdm1d是参与小鼠胚胎发育过程的一个重要基因,在小鼠胚胎早期脑的发育、骨骼生长、多组织形成等过程中可能起重要的作用。初步分析Jhdm1d基因的表达谱为该基因在功能方面的研究提供一定的理论依据。 【英文摘要】The Jhdm1d gene which is named jumonjiC domain-containing of the histone demethylase 1 homologues D is located on mouse chromosome 6. It encodes a 940–amino-acid histone demethylase protein that belongs to jumonji family containing the JmjC domain. JmjC domain is considered to be a common motif in a novel class of histone demethylase, and can demethylate di-methylated Lysine 9 and 27 on Histone H3 (H3K9me2, H3K27me2). Some proteins in the jumonji family may regulate chromatin and transcription inhibition. In order to explore the correlations between the gene expression and tissue development during mouse embryonic development, we made an initiatory analysis to study the expression of the Jhdm1d gene in mRNA level at diverse developmental embryo stages, and analyzed the function of the histone demethylase during mouse development to establish the theory foundation.We analyzed the expression pattern of the Jhdm1d gene using in situ hybridization with mouse embryo as the experimental materials. Whole mount in situ