scholarly journals Visualizing the activity of Escherichia coli divergent promoters and probing their dependence on superhelical density using dual-colour fluorescent reporter vector

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Irina S. Masulis ◽  
Zaira Sh. Babaeva ◽  
Sergey V. Chernyshov ◽  
Olga N. Ozoline
Keyword(s):  
Escherichia Coli ◽  
Fluorescent Reporter ◽  
Divergent Promoters ◽  
Reporter Vector ◽  
Superhelical Density ◽  
Dual Colour

scholarly journals Construction of a Reporter Vector for the Analysis of Bifidobacterium longum Promoters

2006 ◽  
Vol 72 (11) ◽  
pp. 7401-7405 ◽  
Author(s):  
A. Klijn ◽  
D. Moine ◽  
M. Delley ◽  
A. Mercenier ◽  
F. Arigoni ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bifidobacterium Longum ◽  
Cryptic Plasmid ◽  
Reporter Plasmid ◽  
Microarray Experiments ◽  
Reporter Vector

ABSTRACT In order to initiate studies on promoter activities in Bifidobacterium longum and to independently confirm transcriptional data generated by microarray experiments, we have constructed a versatile reporter plasmid based on a B. longum cryptic plasmid and the Escherichia coli gusA gene. The resulting plasmid, pMDY23, has been tested using three B. longum promoters.

scholarly journals Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST

2020 ◽  
Author(s):  
Yankel Chekli ◽  
Caroline Peron-Cane ◽  
Dario Dell’Arciprete ◽  
Jean-François Allemand ◽  
Chenge Li ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Cell Surface ◽  
Bacterial Cell ◽  
Surface Proteins ◽  
Cellular Functions ◽  
Reporter Protein ◽  
Cell Surface Proteins ◽  
Fluorescent Reporter ◽  
Bacterial Proteins

AbstractBacterial proteins exported to the cell surface play key cellular functions. However, despite the interest to study the localization of surface proteins such as adhesins, transporters or hydrolases, monitoring their dynamics in live imaging remains challenging, due to the limited availability of fluorescent probes remaining functional after secretion. In this work, we used the Escherichia coli intimin and the Listeria monocytogenes InlB invasin as surface exposed scaffolds fused with the recently developed chemogenetic fluorescent reporter protein FAST. Using both membrane permeant (HBR-3,5DM) and non-permeant (HBRAA-3E) fluorogens that fluoresce upon binding to FAST, we demonstrated that fully functional FAST can be exposed at the cell surface and specifically tagged on the external side of the bacterial envelop in both diderm and monoderm bacteria. Our work opens new avenues to study of the organization and dynamics of the bacterial cell surface proteins.

An adenovirus-based fluorescent reporter vector to identify and isolate HIV-infected cells

2002 ◽  
Vol 99 (1-2) ◽  
pp. 9-21 ◽  
Author(s):  
Larry Richman ◽  
Pascal R.A Meylan ◽  
Miguel Munoz ◽  
Stéphane Pinaud ◽  
Jovan Mirkovitch
Keyword(s):  
Fluorescent Reporter ◽  
Reporter Vector ◽  
Infected Cells

scholarly journals Promoter Interference in a Bacteriophage Lambda Control Region: Effects of a Range of Interpromoter Distances

2000 ◽  
Vol 182 (1) ◽  
pp. 216-220 ◽  
Author(s):  
Michael G. Strainic ◽  
Jennifer J. Sullivan ◽  
Julio Collado-Vides ◽  
Pieter L. deHaseth
Keyword(s):  
Escherichia Coli ◽  
Complex Formation ◽  
Rna Polymerase ◽  
Control Region ◽  
Bacteriophage Lambda ◽  
Detailed Characterization ◽  
Phosphodiester Bonds ◽  
Divergent Promoters ◽  
Polymerase Binding

ABSTRACT The pR and pRM promoters of bacteriophage lambda direct transcription in divergent directions from start sites separated by 83 phosphodiester bonds. We had previously shown that the presence of an RNA polymerase at pR interfered with open complex formation at pRM and that this effect was alleviated by the deletion of 10 bp between the two promoters. Here we present a detailed characterization of the dependence of the interference on the interpromoter distance. It was found that the reduced interference between the two promoters is unique to the 10-bp deletion. The relief of interference was demonstrated to be due to the facilitation of a step subsequent to RNA polymerase binding to the pRM promoter. A model to explain these observations is proposed. A search of known Escherichia coli promoters identified three pairs of divergent promoters with similar separations to those investigated here.

scholarly journals Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yankel Chekli ◽  
Caroline Peron-Cane ◽  
Dario Dell’Arciprete ◽  
Jean-François Allemand ◽  
Chenge Li ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Cell Surface ◽  
Bacterial Cell ◽  
Surface Proteins ◽  
Cellular Functions ◽  
Reporter Protein ◽  
Cell Surface Proteins ◽  
Fluorescent Reporter ◽  
Bacterial Proteins

Abstract Bacterial proteins exported to the cell surface play key cellular functions. However, despite the interest to study the localisation of surface proteins such as adhesins, transporters or hydrolases, monitoring their dynamics in live imaging remains challenging, due to the limited availability of fluorescent probes remaining functional after secretion. In this work, we used the Escherichia coli intimin and the Listeria monocytogenes InlB invasin as surface exposed scaffolds fused with the recently developed chemogenetic fluorescent reporter protein FAST. Using both membrane permeant (HBR-3,5DM) and non-permeant (HBRAA-3E) fluorogens that fluoresce upon binding to FAST, we demonstrated that fully functional FAST can be exposed at the cell surface and used to specifically tag the external side of the bacterial envelop in both diderm and monoderm bacteria. Our work opens new avenues to study the organization and dynamics of the bacterial cell surface proteins.

scholarly journals The Escherichia coli CpxA-CpxR Envelope Stress Response System Regulates Expression of the Porins OmpF and OmpC

2005 ◽  
Vol 187 (16) ◽  
pp. 5723-5731 ◽  
Author(s):  
Eric Batchelor ◽  
Don Walthers ◽  
Linda J. Kenney ◽  
Mark Goulian
Keyword(s):  
Escherichia Coli ◽  
Stress Response ◽  
Response Regulator ◽  
Gene Encoding ◽  
Fluorescent Reporter ◽  
Response System ◽  
Dnase I Footprinting ◽  
Envelope Stress ◽  
Two Component ◽  
Stress Response System

ABSTRACT We performed transposon mutagenesis of a two-color fluorescent reporter strain to identify new regulators of the porin genes ompF and ompC in Escherichia coli. Screening of colonies by fluorescence microscopy revealed numerous mutants that exhibited interesting patterns of porin expression. One mutant harbored an insertion in the gene encoding the histidine kinase CpxA, the sensor for a two-component signaling system that responds to envelope stress. The cpxA mutant exhibited increased transcription of ompC and a very strong decrease in transcription of ompF under conditions in which acetyl phosphate levels were high. Subsequent genetic analysis revealed that this phenotype is dependent on phosphorylation of the response regulator CpxR and that activation of CpxA in wild-type cells results in similar regulation of porin expression. Using DNase I footprinting, we demonstrated that CpxR binds upstream of both the ompF and ompC promoters. It thus appears that two distinct two-component systems, CpxA-CpxR and EnvZ-OmpR, converge at the porin promoters. Within the context of envelope stress, outer membrane beta-barrel proteins have generally been associated with the sigma E pathway. However, at least for the classical porins OmpF and OmpC, our results show that the Cpx envelope stress response system plays a role in regulating their expression.

scholarly journals Engineering a Circular Riboregulator in Escherichia coli

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
William Rostain ◽  
Shensi Shen ◽  
Teresa Cordero ◽  
Guillermo Rodrigo ◽  
Alfonso Jaramillo
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Messenger Rna ◽  
Noncoding Rna ◽  
Circular Rna ◽  
Circular Rnas ◽  
Fluorescent Reporter ◽  
Small Noncoding Rna ◽  
E Coli ◽  
Group I

RNAs of different shapes and sizes, natural or synthetic, can regulate gene expression in prokaryotes and eukaryotes. Circular RNAs have recently appeared to be more widespread than previously thought, but their role in prokaryotes remains elusive. Here, by inserting a riboregulatory sequence within a group I permuted intron-exon ribozyme, we created a small noncoding RNA that self-splices to produce a circular riboregulator in Escherichia coli. We showed that the resulting riboregulator can trans-activate gene expression by interacting with a cis-repressed messenger RNA. We characterized the system with a fluorescent reporter and with an antibiotic resistance marker, and we modeled this novel posttranscriptional mechanism. This first reported example of a circular RNA regulating gene expression in E. coli adds to an increasing repertoire of RNA synthetic biology parts, and it highlights that topological molecules can play a role in the case of prokaryotic regulation.

scholarly journals The expression of the Escherichia coli fis gene is strongly dependent on the superhelical density of DNA

2000 ◽  
Vol 38 (1) ◽  
pp. 167-175 ◽  
Author(s):  
Robert Schneider ◽  
Andrew Travers ◽  
Georgi Muskhelishvili
Keyword(s):  
Escherichia Coli ◽  
Superhelical Density
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