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

Studies on physiological roles of galactolipase in the chloroplasts and cloning of a cDNA encoding galactolipase from Phaseolus vulgaris L.

叶绿体中半乳糖脂肪酶的生理作用研究及菜豆半乳糖脂肪酶cDNA的克隆。

基本信息

批准号：
04640633
负责人：
SAKAI Takeshi
金额：
$ 0.26万
依托单位：
Hokkaido Tokai 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：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

项目摘要

In this research project, we studied physiological roles of galactolipase in the stress-induced lipid metabolism, immunological properties of galactolipase, and in addition, we tried to isolate a cDNA encoding galactolipase from kidney bean leaves. The results obtained are as follows :1. The metabolic changes from galactolipids to triacylglycerols are results of the enhancement of galactolipase activity in chloroplasts. The lipid changes was shown to occur not only in spinach but also occurred in leaves subjected to SO_2 treatment, water deficiency, and protoplast isolation.2. At least two types of galactolipase, membrane-bound in chloroplasts and soluble, have been reported in kidney bean and runner bean leaves and prepared rabbit antiserum against this enzyme. The antiserum completely inhibited galactolipase activities not only in the chloroplast fractions but also in the soluble ones. Western blot of both solubilized chloroplast proteins and soluble proteins showed single stained band, suggesting the absence of immunologically different isozymes in both fractions.3. A cDNA library was constructed from poly (A) ^+ RNA which was isolated from younger leaves of kidney bean and purified by oligo (dT)-cellulose column. Early screening of the library using antiserum against galactolipase was unsuccessful. After the solution of the sevsral problems and the reconstruction of the library, we now continue to isolate a cDNA clone encoding galactolipase.

在本研究项目中，我们研究了半乳糖酶在胁迫诱导的脂代谢中的生理作用，半乳糖酶的免疫学性质，并试图从菜豆叶片中分离到编码半乳糖酶的基因。研究结果如下：1.半乳糖脂向三酰甘油的代谢变化是叶绿体半乳糖脂酶活性增强的结果。结果表明，不仅菠菜体内的脂类物质发生了变化，SO2处理、水分亏缺和原生质体分离后的叶片也发生了脂类变化。据报道，菜豆和菜豆叶片中至少存在两种类型的半乳糖脂酶，一种是膜结合的，另一种是可溶性的，并制备了抗该酶的兔抗血清。抗血清不仅能完全抑制叶绿体部分的半乳糖脂酶活性，而且能完全抑制叶绿体部分的半乳糖脂酶活性。可溶性叶绿体蛋白和可溶性蛋白的Western印迹均显示单一染色条带，表明这两个组分中没有免疫差异的同工酶。从菜豆幼叶中提取Poly(A)~+RNA，经寡聚(DT)-纤维素柱层析纯化，构建了菜豆Poly(A)~+RNA文库。用半乳糖脂酶抗血清早期筛选文库未获成功。在解决了几个问题并重建了文库后，我们现在继续分离编码半乳脂酶的cDNA克隆。