2020BIO-PROTOCOLOpen access

Measurement of the Promoter Activity in Escherichia coli by Using a Luciferase Reporter

Yuki Yamanaka, Hiroki Watanabe, Erika Yamauchi, Yukari Miyake, Kaneyoshi Yamamoto

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Abstract

The reporter system is widely used technique for measuring promoter activity in bacterial cells.Until now, a number of reporter system have been developed, but the bioluminescent reporter constructed from the bacterial luciferase genes is one of the useful systems for measuring in vivo dynamics of gene expression.The introduced bioluciferase lux reporter enables easy, fast, and sensitive measurement of the promoter activity without cell lysis because the substrates of bioluminescent reaction are synthesized inside the bacterial cell, thereby allowing low-cost experiments.This protocol describes a high throughput technique to measure the promoter activity in Escherichia coli K-12 using the lux reporter system.

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What this paper is about

The reporter system is widely used technique for measuring promoter activity in bacterial cells.Until now, a number of reporter system have been developed, but the bioluminescent reporter constructed from the bacterial luciferase genes is one of the useful systems for measuring in vivo dynamics of gene expression.The introduced bioluciferase lux reporter enables easy, fast, and sensitive measurement of the promoter activity without cell lysis because the substrates of bioluminescent reaction are synthesized inside the bacterial cell, thereby allowing low-cost experiments.This protocol describes a high throughput technique to measure the promoter activity in Escherichia coli K-12 using the lux reporter system.

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Available abstract

The reporter system is widely used technique for measuring promoter activity in bacterial cells.Until now, a number of reporter system have been developed, but the bioluminescent reporter constructed from the bacterial luciferase genes is one of the useful systems for measuring in vivo dynamics of gene expression.The introduced bioluciferase lux reporter enables easy, fast, and sensitive measurement of the promoter activity without cell lysis because the substrates of bioluminescent reaction are synthesized inside the bacterial cell, thereby allowing low-cost experiments.This protocol describes a high throughput technique to measure the promoter activity in Escherichia coli K-12 using the lux reporter system.

Key concepts: Bioreporter, Luciferase, Reporter gene, Bioluminescence, Escherichia coli, Luciferases, Lysis, Chemistry

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