scholarly journals Development and Characterization of a Xylose-Inducible Gene Expression System for Clostridium perfringens

2011 ◽  
Vol 77 (23) ◽  
pp. 8439-8441 ◽  
Author(s):  
Hirofumi Nariya ◽  
Shigeru Miyata ◽  
Tomomi Kuwahara ◽  
Akinobu Okabe
Keyword(s):  
Gene Expression ◽  
Clostridium Perfringens ◽  
Expression System ◽  
Xylose Isomerase ◽  
Inducible Gene Expression ◽  
Content Type ◽  
Regulated Expression ◽  
Operator Sequence ◽  
Inducible Gene

ABSTRACTA xylose-inducible gene expression vector forClostridium perfringenswas developed. Plasmid pXCH contains a chromosomal region fromClostridium difficile(xylR-PxylB):xylR, encoding the xylose repressor,xylO, thexyloperator sequence, and PxylB, the divergent promoter upstream ofxylBAencoding xylulo kinase and xylose isomerase. pXCH allows tightly regulated expression of the chloramphenicol acetyltransferase reporter and the α-toxin genes in response to the inducer concentration. Thus, pXCH could constitute a new valuable genetic tool for study ofC. perfringens.

scholarly journals Cumate-Inducible Gene Expression System for Sphingomonads and Other Alphaproteobacteria

2013 ◽  
Vol 79 (21) ◽  
pp. 6795-6802 ◽  
Author(s):  
Andreas Kaczmarczyk ◽  
Julia A. Vorholt ◽  
Anne Francez-Charlot
Keyword(s):  
Gene Expression ◽  
Caulobacter Crescentus ◽  
Paracoccus Denitrificans ◽  
Expression System ◽  
Model Organisms ◽  
Inducible Gene Expression ◽  
Gene Expression System ◽  
Content Type ◽  
Wide Range ◽  
Inducible Gene

ABSTRACTTunable promoters represent a pivotal genetic tool for a wide range of applications. Here we present such a system for sphingomonads, a phylogenetically diverse group of bacteria that have gained much interest for their potential in bioremediation and their use in industry and for which no dedicated inducible gene expression system has been described so far. A strong, constitutive synthetic promoter was first identified through a genetic screen and subsequently combined with the repressor and the operator sites of thePseudomonas putidaF1cym/cmtsystem. The resulting promoter, termed PQ5, responds rapidly to the inducer cumate and shows a maximal induction ratio of 2 to 3 orders of magnitude in the different sphingomonads tested. Moreover, it was also functional in otherAlphaproteobacteria, such as the model organismsCaulobacter crescentus,Paracoccus denitrificans, andMethylobacterium extorquens. In the noninduced state, expression from PQ5is low enough to allow gene depletion analysis, as demonstrated with the essential genephyPofSphingomonassp. strain Fr1. A set of PQ5-based plasmids has been constructed allowing fusions to affinity tags or fluorescent proteins.

scholarly journals Construction and Characterization of a Gradually Inducible Expression Vector for Halobacterium salinarum, Based on thekdpPromoter

2012 ◽  
Vol 78 (7) ◽  
pp. 2100-2105 ◽  
Author(s):  
Dorthe Kixmüller ◽  
Jörg-Christian Greie
Keyword(s):  
Gene Expression ◽  
Expression Vector ◽  
Expression System ◽  
Expression Patterns ◽  
Inducible Expression ◽  
Halobacterium Salinarum ◽  
Expression Vectors ◽  
Content Type ◽  
Inducible Expression System

ABSTRACTGradually inducible expression vectors which are governed by variations of growth conditions are powerful tools for gene expression of conditionally lethal mutants. Furthermore, controlled expression allows monitoring of overproduction of proteins at various stages in their expressing hosts. ForHalobacterium salinarum, which is often used as a paradigm for halophilic archaea, such an inducible expression system is not available to date. Here we show that thekdppromoter (Pkdp), which facilitates gene expression upon K+limitation, can be used to establish such a system for molecular applications. Pkdpfeatures a rather high expression rate, with an approximately 50-fold increase that can be easily varied by K+concentrations in the growth medium. Besides the construction of an expression vector, our work describes the characterization of expression patterns and, thus, offers a gradually inducible expression system to the scientific community.

scholarly journals Implementation of the agmatine-controlled expression system for inducible gene expression in Lactococcus lactis

2015 ◽  
Vol 14 (1) ◽  
Author(s):  
Daniel M. Linares ◽  
Patricia Alvarez-Sieiro ◽  
Beatriz del Rio ◽  
Victor Ladero ◽  
Begoña Redruello ◽  
...  
Keyword(s):  
Gene Expression ◽  
Lactococcus Lactis ◽  
Expression System ◽  
Inducible Gene Expression ◽  
Inducible Gene

Heat-Inducible Gene Expression System by Applying Alternating Magnetic Field to Magnetic Nanoparticles

2013 ◽  
Vol 3 (5) ◽  
pp. 273-279 ◽  
Author(s):  
Masaki Yamaguchi ◽  
Akira Ito ◽  
Akihiko Ono ◽  
Yoshinori Kawabe ◽  
Masamichi Kamihira
Keyword(s):  
Gene Expression ◽  
Magnetic Field ◽  
Magnetic Nanoparticles ◽  
Expression System ◽  
Alternating Magnetic Field ◽  
Inducible Gene Expression ◽  
Gene Expression System ◽  
Inducible Gene

scholarly journals An Archaeal Fluoride-Responsive Riboswitch Provides an Inducible Expression System for Hyperthermophiles

2018 ◽  
Vol 84 (7) ◽  
Author(s):  
Michael Clayton Speed ◽  
Brett W. Burkhart ◽  
Jonathan W. Picking ◽  
Thomas J. Santangelo
Keyword(s):  
Gene Expression ◽  
Transcription Initiation ◽  
Expression System ◽  
Regulatory Control ◽  
Cell Physiology ◽  
Content Type ◽  
Thermococcus Kodakarensis ◽  
Export Pathway ◽  
Regulated Expression ◽  
Regulated Gene Expression

ABSTRACT Robust genetic systems for the hyperthermophilic Thermococcales have facilitated the overexpression of native genes, enabled the addition of sequences encoding secretion signals, epitope, and affinity tags to coding regions, and aided the introduction of sequences encoding new proteins in these fast-growing fermentative heterotrophs. However, tightly controlled and easily manipulated systems facilitating regulated gene expression are limited for these hosts. Here, we describe an alternative method for regulatory control reliant on a cis -encoded functional riboswitch in the model archaeon Thermococcus kodakarensis . Despite the hyperthermophilic growth temperatures, the proposed structure of the riboswitch conforms to a fluoride-responsive riboswitch encoded in many bacteria and similarly functions to regulate a component-conserved fluoride export pathway. Deleting components of the fluoride export pathway generates T. kodakarensis strains with increased fluoride sensitivity. The mechanism underlying regulated expression suggested that the riboswitch-encoding sequences could be utilized as a tunable expression cassette. When appended to a reporter gene, the riboswitch-mediated control system provides fluoride-dependent tunable regulatory potential, offering an alternative system for regulating gene expression. Riboswitch-regulated expression is thus ubiquitous in extant life and can be exploited to generate regulated expression systems for hyperthermophiles. IMPORTANCE Gene expression is controlled by a myriad of interconnected mechanisms that interpret metabolic states and environmental cues to balance cell physiology. Transcription regulation in Archaea is known to employ both typical repressors-operators and transcription activators to regulate transcription initiation in addition to the regulation afforded by chromatin structure. It was perhaps surprising that the presumed ancient mechanism of riboswitch-mediated regulation is found in Bacteria and Eukarya , but seemingly absent in Archaea . We demonstrate here that a fluoride-responsive riboswitch functions to regulate a detoxification pathway in the hyperthermophilic archaeon Thermococcus kodakarensis . The results obtained define a universal role for riboswitch-mediated regulation, adumbrate the presence of several riboswitch-regulated genes in Thermococcus kodakarensis , demonstrate the utility of RNA-based regulation at high temperatures, and provide a novel riboswitch-regulated expression system to employ in hyperthermophiles.

scholarly journals A tunable anthranilate-inducible gene expression system for Pseudomonas species

2020 ◽  
Vol 105 (1) ◽  
pp. 247-258
Author(s):  
Lena Hoffmann ◽  
Michael-Frederick Sugue ◽  
Thomas Brüser
Keyword(s):  
Gene Expression ◽  
Expression System ◽  
Ant System ◽  
Plant Host ◽  
Inducible Gene Expression ◽  
Gene Expression System ◽  
Expression Levels ◽  
Key Points ◽  
Wide Range ◽  
Inducible Gene

Abstract Pseudomonads are among the most common bacteria in soils, limnic ecosystems, and human, animal, or plant host environments, including intensively studied species such as Pseudomonas aeruginosa, P. putida, or P. fluorescens. Various gene expression systems are established for some species, but there is still a need for a simple system that is suitable for a wide range of pseudomonads and that can be used for physiological applications, i.e., with a tuning capacity at lower expression levels. Here, we report the establishment of the anthranilate-dependent PantA promoter for tunable gene expression in pseudomonads. During studies on P. fluorescens, we constructed an anthranilate-inducible AntR/PantA-based expression system, named pUCP20-ANT, and used GFP as reporter to analyze gene expression. This system was compared with the rhamnose-inducible RhaSR/PrhaB-based expression system in an otherwise identical vector background. While the rhamnose-inducible system did not respond to lower inducer concentrations and always reached high levels over time when induced, expression levels of the pUCP20-ANT system could be adjusted to a range of distinct lower or higher levels by variation of anthranilate concentrations in the medium. Importantly, the anthranilate-inducible expression system worked also in strains of P. aeruginosa and P. putida and therefore will be most likely useful for physiological and biotechnological purposes in a wide range of pseudomonads. Key points • We established an anthranilate-inducible gene expression system for pseudomonads. • This system permits tuning of gene expression in a wide range of pseudomonads. • It will be very useful for physiological and biotechnological applications.

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