SEPARATION AND CHARACTERIZATION OF ARABIDOPSIS-THALIANA PROTEINS BY 2-DIMENSIONAL GEL-ELECTROPHORESIS
SEPARATION AND CHARACTERIZATION OF ARABIDOPSIS-THALIANA PROTEINS BY 2-DIMENSIONAL GEL-ELECTROPHORESIS
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DOI：
10.1002/elps.1150160169
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发表时间：
1995-03-01
期刊：
影响因子：
2.9
通讯作者：
TSUGITA, A
中科院分区：
文献类型：
--
作者：
KAMO, M;KAWAKAMI, T;TSUGITA, A
从拟南芥（Arabidopsis thaliana）5种组织（叶、茎、根、种子和愈伤组织）中提取蛋白质，采用双向凝胶电泳（2-DE）进行分离。双向电泳采用固定化pH梯度法（IPG）进行第一维双向电泳，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）进行第二维双向电泳。借助共迁移的五个标记蛋白，每个组织的2-DE凝胶的图案由计算机图形地组合成一个单一的合成图像，用于整合的拟南芥蛋白质点。共获得4763张蛋白质斑点图，分别用分子量和等电点表征。用Edman降解法分析了101个蛋白质点的N端部分序列。对57个蛋白质进行了部分测序，46个蛋白质似乎具有封闭的N-末端。用肼蒸气对14种蛋白质进行了去封闭，发现其中两种是焦谷氨酰封闭的N-末端。本研究发现了47个新蛋白。
Arabidopsis (Arabidopsis thaliana) proteins were isolated from five tissues (leaf, stem, root, seed and callus), and separated by two-dimensional gel electrophoresis (2-DE). 2-DE was carried out by immobilized pH gradient (IPG) in the first dimension, and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension. With the aid of comigrated five-marker proteins, the patterns of 2-DE gels for each tissue were graphically combined by a computer into a single synthetic image for the integrated Arabidopsis protein spots. The protein spot images, altogether 4763, were characterized by both molecular mass and isoelectric point. Partial amino(N)-terminal sequences of 101 protein spots were analyzed by Edman degradation. Fifty seven proteins were partially sequenced and 46 proteins appeared to have blocked N-termini. Deblocking by hydrazine vapor was carried out on 14 proteins and two of them were found to be pyroglutamyl-blocked N-termini. Forty seven new proteins were found by the present investigation.