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Development of a new Chromosome-Identification Method and Gene-Mapping of Xenopus Laevis Chromosome.

非洲爪蟾染色体新染色体鉴定方法和基因图谱的开发。

基本信息

批准号：
01540529
负责人：
KONDO Toshihiko
金额：
$ 1.09万
依托单位：
Gunma University
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-01540529/
关键词：
Chromosome Chromosome map Gene mapping Xenopus laevis Sex determination mechanism Metamorphosis 性決定遺伝子

项目摘要

In order to identify Xenopus chromosomes and to make a gene-map of Xenopus chromosome, we tried to develop some methods, and cloning many chromosome specific genes, hormone genes, sex specific genes (or sex determining genes), and metamorphosis (or programmed cell death) marker genes.We developed ; 1. a highly sensitive nucleic acid detection method and gene-mapping method using chain reaction of fluorescence amplifiers, 2. a rapid isolation method for highly purified plasmid DNA, 3. a high resolution HPLC system using strong basic anion exchange resin (QA column), and 4. a DNA-immobilized anchor (DNA is enzymatically solidified by DNA ligase). By combination of these methods, we established ; 5. a new solid-phase cloning system, and 6. a hybrid selection method. According to use these method, we cloned ; 7. female specific DNAs (DNA markers of sex chromosome of Xenopus laevis, and 8. collagenase gene (a marker gene of metamorphosis in the amphibia). And, we are further cloning many chromosome specific gene markers and other marker genes. In addition to this research, we found an epidermal factor which stimulates collagenase production by fibroblasts in a reconstituted skin model, and clarified the hormonogenetic donor sites of thyroid hormone in thyroglobulin.

为了鉴定非洲爪哇的染色体和绘制非洲爪哇染色体的基因图谱，我们尝试了一些方法，克隆了许多染色体特异基因、激素基因、性别决定基因(或性别决定基因)和变态(或程序性细胞死亡)标记基因。我们建立了一种高灵敏的核酸检测方法和基于荧光扩增链式反应的基因定位方法，2.一种高纯度的质粒DNA的快速提取方法，3.一种使用强碱性阴离子交换树脂(QA柱)的高分辨率高效液相色谱系统，以及4.DNA固定化锚(DNA由DNA连接酶催化固化)。通过这些方法的结合，我们建立了：5.新的固相克隆系统，6.杂交选择方法。根据这些方法，我们克隆了：7.雌性特有的DNA(非洲爪哇性染色体的DNA标记)；8.胶原酶基因(两栖动物变态的标记基因)。此外，我们还在进一步克隆许多染色体特异的基因标记和其他标记基因。除了这项研究，我们还在重组皮肤模型中发现了一种刺激成纤维细胞产生胶原酶的表皮因子，并阐明了甲状腺球蛋白中甲状腺激素的激素供体部位。