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全组织标本中lacZ融合蛋白的β-半乳糖苷酶染色

β-Galactosidase staining of lacZ fusion proteins in whole tissue preparations.

作者信息

Cooper Margaret A, Zhou Renping

机构信息

Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.

出版信息

Methods Mol Biol. 2013;1018:189-97. doi: 10.1007/978-1-62703-444-9_18.

Abstract

The lacZ gene product, β-galactosidase, has classically been used as a reporter of gene expression. β-Galactosidase activity can be detected using a chromogenic substrate, X-gal, which leaves an intense blue precipitate when cleaved by the enzyme. Insertion of the lacZ coding DNA targeted into a specific gene creates a β-galactosidase-tagged fusion protein that is expressed under the endogenous promoter. Analysis of the hybrid protein takes advantage of the chromogenic detection system, as the distribution and relative abundance of the expressed protein can be efficiently visualized.

摘要

LacZ基因产物β-半乳糖苷酶一直被经典地用作基因表达的报告分子。β-半乳糖苷酶活性可以使用生色底物X-gal进行检测,当该底物被酶切割时会留下强烈的蓝色沉淀。将lacZ编码DNA插入到特定基因中会产生一个β-半乳糖苷酶标记的融合蛋白,该融合蛋白在内源启动子的控制下表达。对杂交蛋白的分析利用了生色检测系统,因为所表达蛋白的分布和相对丰度可以有效地可视化。

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