Roles of neurotrophic factors derived from astrocyte in neuronal development

(1)人gmf - γ cDNA的克隆我们从人胎脑cDNA文库中分离到一段人gmf - γ cDNA。其中一个克隆包含一个573个核苷酸长的插入和一个426 bp的开放阅读框，编码一个推断的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插入pAED4表达载体，导入大肠杆菌BL21（DE3）。将表达的蛋白纯化并进行抗体生产。(3)抗人gmf - β和gmf - γ抗体的制备以重组人gmf - β和gmf - γ作为免疫原制备多克隆抗体。各抗血清分别识别人gmf - β和gmf - γ，但对大鼠gmf - β和gmf - γ均无反应。(4)大鼠gmf - γ cDNA的克隆我们从大鼠肺动脉cDNA文库中分离到一段大鼠gmf - γ cDNA。其中一个克隆包含一个553个核苷酸长的插入和一个426 bp的开放阅读框，编码一个推断的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插入pAED4表达载体，导入大肠杆菌BL21（DE3）。将表达的蛋白纯化并进行抗体生产。(6)抗大鼠gmf - β和gmf - γ抗体的制备采用与人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。

(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.