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从聚丙烯酰胺凝胶电泳电转移至固定化基质上的蛋白质的微量测序:应用于一种不溶性蛋白质

Microsequencing of proteins electrotransferred onto immobilizing matrices from polyacrylamide gel electrophoresis: application to an insoluble protein.

作者信息

Hirano H, Watanabe T

机构信息

National Institute of Agrobiological Resources, Tsukuba, Japan.

出版信息

Electrophoresis. 1990 Jul;11(7):573-80. doi: 10.1002/elps.1150110708.

Abstract

Semidry electroblotting is convenient and allows a rapid and efficient protein transfer from one- or two-dimensional polyacrylamide gels onto sequencer stable supports for protein microsequence analysis in a gas-phase sequencer. Using this technique, we determined the amino acid sequences of the basic 7S globulin (Bg), an insoluble protein present in soybean seeds. Based on sequence determination, the cDNA-encoding Bg could be easily cloned and characterized.

摘要

半干电印迹法操作简便,能快速高效地将蛋白质从一维或二维聚丙烯酰胺凝胶转移至测序稳定支持物上,以便在气相测序仪中进行蛋白质微序列分析。运用该技术,我们测定了大豆种子中存在的一种不溶性蛋白质——碱性7S球蛋白(Bg)的氨基酸序列。基于序列测定结果,编码Bg的cDNA能够轻松地被克隆和鉴定。

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