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Thermogenetic tools to monitor temperature-dependent gene expression in bacteria

Paper ID Volume ID Publish Year Pages File Format Full-Text
23538 43449 2012 9 PDF Available
Title
Thermogenetic tools to monitor temperature-dependent gene expression in bacteria
Abstract

Free-living bacteria constantly monitor their ambient temperature. Drastic deviations elicit immediate protective responses known as cold shock or heat shock response. Many mammalian pathogens use temperature surveillance systems to recognize the successful invasion of a host by its body temperature, usually 37 °C. Translation of temperature-responsive genes can be modulated by RNA thermometers (RNATs). RNATs form complex structures primarily in the 5′-untranslated region of their transcripts. Most RNATs block the ribosome binding site at low temperatures. Translation is induced at increasing temperature by melting of the RNA structure. The analysis of such temperature-dependent RNA elements calls for adequate test systems that function in the appropriate temperature range. Here, we summarize previously established reporter gene systems based on the classical β-galactosidase LacZ, the heat-stable β-galactosidase BgaB and the green fluorescent protein GFP. We validate these systems by testing known RNATs and describe the construction and application of an optimized bgaB system. Finally, two novel RNA thermometer candidates from Escherichia coli and Salmonella will be presented.

► Several reporter gene systems can be used to analyze RNA thermometers in vivo. ► Both inducible as well as constitutively regulated systems are applicable. ► GFP and the β-galactosidases LacZ and BgaB (thermostable) are suitable reporters. ► Besides moderate heat shock, LacZ is optimal for analyzing cold shock regulation. ► Novel RNA thermometer candidates were found upstream of the htrA gene from Salmonella and Escherichia coli.

Keywords
bp(s), base pair(s); Csp, cold shock protein; fourU, four uridines; Hsp, heat shock protein; nt, nucleotides; RBS, ribosomal binding site; RNAT, RNA thermometer; ROSE, repression of heat shock gene expression; SD, Shine-Dalgarno; TSS, transcriptional star
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 160, Issues 1–2, 31 July 2012, Pages 55–63
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us