Promoter analysis by saturation mutagenesis

被引：8

作者：

Baliga N.S. ^{[1
]}

机构：

[1] Institute for Systems Biology, Seattle, WA 98105

来源：

Biological Procedures Online | 2001年 / 3卷 / 1期

关键词：

Mutagenesis; Polymerase chain reaction; Promoter regions (genetics);

D O I：

10.1251/bpo24

中图分类号：

学科分类号：

摘要：

Gene expression and regulation are mediated by DNA sequences, in most instances, directly upstream to the coding sequences by recruiting transcription factors, regulators, and a RNA polymerase in a spatially defined fashion. Few nucleotides within a promoter make contact with the bound proteins. The minimal set of nucleotides that can recruit a protein factor is called a cis-acting element. This article addresses a powerful mutagenesis strategy that can be employed to define cis-acting elements at a molecular level. Technical details including primer design, saturation mutagenesis, construction of promoter libraries, phenotypic analysis, data analysis, and interpretation are discussed. © 2001 Biological Procedures Online. All rights reserved.

引用

页码：64 / 69

页数：5

共 13 条

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