Molecular Mechanisms Underlying Differentiation of Microbodies.

微生物分化的分子机制。

基本信息

批准号：
04454017
负责人：
NISHIMURA Mikio
金额：
$ 4.48万
依托单位：
National Institute for Basic Biology
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
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

A cDNA clone for a leaf-peroxisomal enzyme, glycolate oxidase (E C 1.1.3.1), was isolated by an immunochemical method from a cDNA expression library constructed from poly (A)^+ -RNA of green pumpkin cotyledons. Northern blot analysis showed that the probe could hybridizable with a single 1.5-kb species of mRNA from pumpkin cotyledons and that the amount of the hybridizable mRNA increased dramatically during greening of the cotyledons, Immunoblot analysis showed that the amount of glycolate oxidase was low in darkgrown cotyledons and increased during greening of pumpkin cotyledons. This observation indicates that the induction of a leaf peroxisomal enzyme, glycolate oxidase, during greening of the cotyledons is due to an increase in level of the mRNA.After the function transition of glyoxysomes to leaf peroxisomes during the greening of pumpkin cotyledons, the reverse microbody transition of leaf peroxisomes to glyoxysomes occurs during senescence. Immunocytochemical labeling with protein A-gold was performed to analyze the reverse microbody transition using antibodies against a leaf-peroxisomal enzyme, glycolate oxidase, and ageinst two gloxysomal enzymes, namely, malate synthase and isocitrate lyase. The intensity of labeling for glycolate oxidase decreased in the microbodies during senescence whereas in the case of malate synthase and isocitrate lyase intensities increase strikingly. Double labeling experments with protein A-gold particles of different sizes showed that the leaf-peroxisomal enzymes and the glyoxysomal enzymes coexist in the microbodies of senescing pumpkin cotyledons, indicating that leaf peroxisomal are directly transformed to glyoxysomes during, senescence.

通过免疫化学方法分离出一种由绿色南瓜共叶叶的poly（a）^+ -RNA构成的cDNA表达式文库，通过免疫化学方法分离出用于叶酸氧化酶（E C 1.1.3.1）的叶氧化酶（E C 1.1.3.1）的cDNA克隆。 Northern印迹分析表明，该探针可以与南瓜子叶的单个1.5 kb的mRNA混合杂交，并且在共蛋白酶绿化期间，杂交mRNA的量大大增加，免疫印迹分析表明，在绿色糖果中，糖蜜氧化酶的量较低，而在绿色的cotnings cottin中会增加。该观察结果表明，在子叶绿化期间，叶叶过氧化物酶体氧化酶，糖基氧化酶的诱导是由于mRNA水平的增加所致。糖体的功能过渡到叶氧化叶的叶片过氧化物体的叶片过氧化物体的叶片过氧化物中的绿叶糖体呈叶状性叶片的过渡性，在叶片反向过渡过程中，以叶片的转变为叶片。使用蛋白A-gold进行免疫细胞化学标记，以分析针对叶 - 过氧化物酶体酶，乙醇酸氧化酶的抗体，糖体氧化酶和AgeSTST的两种gloxysomal酶的反向微生物转变，即，麦芽酸合成酶和异位酸酯酶。在衰老过程中，微糖酸氧化酶的标记强度降低，而在苹果酸合酶和异位酸酯裂解酶强度的情况下，显着性会增加。用不同尺寸的蛋白A-gold颗粒进行双重标记实验表明，叶 - 过氧化物酶体酶和乙氧化酶在渗透性南瓜子叶的微生物中共存，这表明叶片过氧化酶体直接转化为透明剂。