Study of the gene expression in development and regulation of methylation of DNA

DNA甲基化发育及调控中基因表达的研究

基本信息

批准号：
01540590
负责人：
YASUGI Sadao
金额：
$ 1.28万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01540590/
关键词：
regulation of gene expression pepsinogen gene embryonic development methylation of DNA

项目摘要

The aim of the present study is to elucidate the regulation mechanisms of genes which show stage-specific and organ-specific expression during animal development. I used for this purpose the embryonic chicken pepsinogen (ECPg) gene which is expressed specifically in the epithelial cells of proventriculus (glandular stomach) during chicken embryonic period.In mouse and human, degree of methylation in DNA of pepsinogen genes has shown to become lower in the organs expressing pepsinogens. So I examined the degree of methylation in ECPg gene obtained from ECPg-expressing and ECPg-non-expressing organs.ECPg gene showed rather wide range polymorphisms when genomic DNA from chicken organs was digested with several restriction enzymes and subjected to Southern hybridization with cDNA of ECPg as a probe. I selected, therefore, the individuals which had the same hybridization pattern for further analysis. DNAs from proventriculus, gizzard, small intestine and muscle showed the same hybridization pattern when they were digested with methylation-sensitive restriction endonucleases such as Hha I or Hpa II. These results indicate that, ECPg expresssion is not regulated by methylation of DNA itself, but by sis- or trans-acting factors in the proventricular epithelial cells.

本研究的目的是阐明基因的调节机制，这些机制在动物发育过程中表现出阶段特异性和器官特异性表达。我用于此目的是在鸡肉胚胎期间特异性地表达在普雷内肠胃（腺体胃）的上皮细胞中的胚胎鸡肉蛋白原（ECPG）基因。在小鼠和人类中，胸肌蛋白原基因DNA的甲基化程度已显示出在表达pepsinogenogenogenogenogenogengans pepsinogenogenogengans基因的DNA程度。因此，我检查了通过表达ECPG和ECPG-NON表达的器官获得的ECPG基因中的甲基化程度。ECPG基因在鸡肉器官的基因组DNA被几种限制酶中消化并与Ecpg of Probe的cdna hybridization一起消化，而鸡肉器官的基因组DNA则显示出相当广泛的多态性。因此，我选择了具有相同杂交模式的个体以进行进一步分析。 ProPentriculus，Gizzard，小肠和肌肉的DNA在用甲基化敏感的限制性核酸内切核酸酶（例如HHA I或HPA II）消化时，它们显示出相同的杂交模式。这些结果表明，ECPG表达不受DNA本身的甲基化的调节，而是由前后上皮细胞中的SIS或反式作用因子调节。