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Roles of neurotrophic factors derived from astrocyte in neuronal development

星形胶质细胞衍生的神经营养因子在神经元发育中的作用

基本信息

批准号：
10670747
负责人：
ASAI Kiyofumi
金额：
$ 1.98万
依托单位：
Nagoya City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670747/
关键词：
Glia Maturation Factor Gene Cloning グリア細胞成長因子 GMF アストロサイト

项目摘要

(1) Cloning of human GMF-gamma cDNAWe isolated a human GMF-gamma cDNA from human fetal brain cDNA library. One of the clones contained a 573 nucleotide long insert and an open reading frame of 426 bp encoding a deduced protein of 142 amino acids.(2) Expression of human GMF-beta and GMF-gammaThe cDNA of human GMF-beta and GMF-gamma were inserted to pAED4 expression vector and introduced to E. coli BL21(DE3). The expressed proteins were purified and subjected to antibody production.(3) Production of anti-human GMF-beta and GMF-gamma antibodiesRecombinant human GMF-beta and GMF-gamma were used as immunogens for the production of polyclonal antibodies. The each anti-serum recognized human GMF-beta and GMF-gamma respectively, but either of them did not react to rat GMF-beta or GMF-gamma.(4) Cloning of rat GMF-gamma cDNAWe isolated a rat GMF-gamma cDNA from rat pulmonary artery cDNA library. One of the clones contained a 553 nucleotide long insert and an open reading frame of 426 bp encoding a deduced protein of 142 amino acids.(5) Expression of rat GMF-beta and GMF-gammaThe cDNA of rat GMF-beta and GMF-gamma were inserted to pAED4 expression vector and introduced to E. coli BL21(DE3). The expressed proteins were purified and subjected to antibody production.(6) Production of anti-rat GMF-beta and GMF-gamma antibodiesAnti-rat GMF-beta and GMF-gamma serum were produced by the same way of human GMF-beta and GMF-gamma. The anti-sera recognized rat GMF-beta and GMF-gamma respectively.

(1)人GMF-γ cDNA的克隆我们从人胎脑cDNA文库中分离到一个人GMF-γ cDNA。其中一个克隆含有573个核苷酸长的插入片段和一个426 bp的开放阅读框，编码142个氨基酸的蛋白质。(2)人GMF-β和GMF-γ的表达将人GMF-β和GMF-γ的cDNA分别插入到pAED 4表达载体中，并导入E. coli BL21（DE3）。纯化表达的蛋白质并进行抗体生产。(3)抗人GMF-β和GMF-γ抗体的制备重组人GMF-β和GMF-γ用作制备多克隆抗体的免疫原。每种抗血清分别识别人GMF-β和GMF-γ，但它们中的任何一种都不与大鼠GMF-β或GMF-γ反应。(4)大鼠GMF-γ cDNA的克隆我们从大鼠肺动脉cDNA文库中分离到一个大鼠GMF-γ cDNA。其中一个克隆含有553个核苷酸长的插入片段和一个426 bp的开放阅读框，编码142个氨基酸的蛋白质。(5)大鼠GMF-β和GMF-γ的表达将大鼠GMF-β和GMF-γ的cDNA分别插入pAED 4表达载体，并导入E. coli BL21（DE3）。纯化表达的蛋白质并进行抗体生产。(6)抗大鼠GMF-β和GMF-γ抗体的制备通过与人GMF-β和GMF-γ相同的方法制备抗大鼠GMF-β和GMF-γ血清。抗血清分别识别大鼠GMF-β和GMF-γ。