权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

The mechanism of differentiation pattern formation in the cellular slime molds as analysed by monoclonal antibodies

单克隆抗体分析细胞粘菌分化模式形成机制

基本信息

批准号：
60480016
负责人：
TAKEUCHI Ikuo
金额：
$ 3.97万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1986
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60480016/
关键词：
Cell differentiation Pattern formation Monoclonal antibodies 細胞性粘菌

项目摘要

1. Two types of cells (prestalk and prespore cells) differentiate in a tissue formed by aggregation of cellular slime mold cells. We tried to obtain monoclonal antibodies specific for each cell type and produced four prespore specific antibodies. Antigens reactive with the antibodies are localized in prespore vacuoles specifically formed in prespore cells and released from the cells upon sporulation to constitute spore coat. By using polyclonal antibodies produced against a molecular species of spore coat proteins, a cDNA library was screened to obtain a prespore specific cDNA clone and a gene for this clone is being isolated. After the gene is obtained, we will examine its structure and regulation of expression together with those of other genes specifically expressed in prespore cells.2. As stalk and prestalk specific proteins, we purified a WGA binding protein (31Kd) and a protein (35Kd) reactive with a cell type-nonspecific monoclonal antibody, but failed to produce monoclonal and polyclonal antibodies for either of them. On the other hand, prestalk specific cDNA clones were used to obtain a large amount of specific proteins, against which we are in the process of producing specific antibodies.3. Some of polyclonal antispore antibodies and prespore specific monoclonal antibodies were found to cross-react with antigens of different species of not only Dictyostelium but also Polysphondylium. As cell differentiation in the latter species has not been well elucidated, we are investigating the process of pattern formation by the use of these antibodies.4. A mutant in which prestalk and prespore cells fail to sort out to produce the two presumptive regions was isolated and we are trying to examine the abnormality of the mutant at protein levels.

1.两种类型的细胞（前柄细胞和前孢子细胞）在由细胞黏菌细胞聚集形成的组织中分化。我们试图获得对每种细胞类型特异的单克隆抗体，并产生了四种前孢子特异性抗体。与抗体反应的抗原定位于前孢子细胞中特异性形成的前孢子液泡中，并在孢子形成时从细胞释放以构成孢子衣。通过使用针对孢子外壳蛋白的分子种类产生的多克隆抗体，筛选cDNA文库以获得前孢子特异性cDNA克隆，并且分离该克隆的基因。获得该基因后，我们将研究其结构和表达调控，以及在前孢子细胞中特异表达的其他基因。作为茎和前茎特异性蛋白，我们纯化了WGA结合蛋白（31Kd）和与细胞类型非特异性单克隆抗体反应的蛋白（35Kd），但未能产生针对它们中任一种的单克隆抗体和多克隆抗体。另一方面，利用前茎特异性cDNA克隆获得了大量的特异性蛋白质，针对这些蛋白质的特异性抗体正在制备过程中.部分抗孢子多克隆抗体和前孢子特异性单克隆抗体不仅与网骨藻属不同种的抗原发生交叉反应，而且与多软骨藻属不同种的抗原发生交叉反应。由于后者物种的细胞分化尚未得到很好的阐明，我们正在研究通过使用这些抗体的模式形成的过程。分离到一个前柄细胞和前孢子细胞不能分离产生两个假定区域的突变体，我们正试图在蛋白质水平上检查突变体的异常。