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等电聚焦和十二烷基硫酸钠凝胶电泳法对醇溶蛋白的序列分析。

Serial analysis of zein by isoelectric focusing and sodium dodecyl sulfate gel electrophoresis.

机构信息

United States Department of Agriculture, Agricultural Research Service, 1102 South Goodwin, Urbana, Illinois 61801.

出版信息

Plant Physiol. 1986 Sep;82(1):196-202. doi: 10.1104/pp.82.1.196.

Abstract

Zein, the major storage protein of maize (Zea mays L.) endosperm, was extracted from a number of inbreds with alcohol plus a reducing agent. Isoelectric focusing (IEF) separated total zeins into 41 components, while sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) separated total zeins into about 15 components. Each procedure gave characteristic patterns of zein bands for a number of maize inbreds. IEF and SDS-PAGE were used serially so that each band separated by IEF could be assayed as an individual SDS-PAGE sample. Some IEF bands revealed only a single band after SDS-PAGE, while others revealed two or more bands. A nomenclature system is presented which integrates the two separation systems with information about chromosome locations of zein genes, maize mutations which affect zein synthesis, and inbred sources for different zeins. SDS-PAGE of zein gives apparent molecular masses which vary widely according to the standards used and the properties of the gels, therefore an artificial nomenclature for identifying zein bands after SDS-PAGE is presented. The new nomenclature provides a flexible system which is useful and can be conveniently used in different laboratories.

摘要

玉米醇溶蛋白是玉米胚乳的主要储存蛋白,可从许多自交系中用酒精加还原剂提取。等电聚焦(IEF)将总醇溶蛋白分离成 41 个组份,而十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)将总醇溶蛋白分离成约 15 个组份。每个程序都为许多玉米自交系产生了醇溶蛋白带的特征模式。IEF 和 SDS-PAGE 连续使用,以便 IEF 分离的每个带都可以作为单个 SDS-PAGE 样品进行分析。一些IEF 带在 SDS-PAGE 后只显示一条带,而其他带则显示两条或更多条带。提出了一种命名系统,该系统将两种分离系统与醇溶蛋白基因的染色体位置、影响醇溶蛋白合成的玉米突变以及不同醇溶蛋白的自交系信息整合在一起。SDS-PAGE 分析醇溶蛋白时,根据所使用的标准和凝胶的性质,其表观分子量差异很大,因此提出了一种用于鉴定 SDS-PAGE 后醇溶蛋白带的人工命名法。新的命名法提供了一个灵活的系统,在不同的实验室中都很有用且方便使用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea6c/1056089/aaad6f89fd35/plntphys00605-0204-a.jpg

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