The identification of bacterial gene expression differences using mRNA-based isothermal subtractive hybridization

1995 ◽  
Vol 41 (2) ◽  
pp. 152-156 ◽  
Author(s):  
Eric A. Utt ◽  
Jeffery P. Brousal ◽  
Lynne C. Kikuta-Oshima ◽  
Frederick D. Quinn
Keyword(s):  
Gene Expression ◽  
Listeria Monocytogenes ◽  
Subtractive Hybridization ◽  
Listeriolysin O ◽  
Bacterial Strains ◽  
Bacterial Gene ◽  
Total Rna ◽  
Bacterial Gene Expression ◽  
Hybridization Probes ◽  
The Difference

We describe a method for isolating and determining differences in gene expression between related bacterial strains. The method is based upon differences in mRNA expression. To demonstrate this procedure, cDNA generated from total RNA of Listeria monocytogenes serotype 1/2a was hybridized to total RNA from a Tn916 mutant of serogroup 1/2a (M3) that was deficient in the production of listeriolysin O, the product of the hly gene. The single-stranded cDNA fragments remaining after hybridization represent the difference in expressed genes between the two strains. These subtraction products were used as hybridization probes to identify the corresponding hly gene in a Southern hybridization.Key words: subtractive hybridization, Listeria monocytogenes, hemolysin, gene expression, isogenic.

scholarly journals Measurement of bacterial gene expression in vivo

2000 ◽  
Vol 355 (1397) ◽  
pp. 601-611 ◽  
Author(s):  
Isabelle Hautefort ◽  
Jay C. D. Hinton
Keyword(s):  
Gene Expression ◽  
Dynamic Environment ◽  
Subtractive Hybridization ◽  
Fluorescence Induction ◽  
Infection Process ◽  
Bacterial Gene ◽  
Technology System ◽  
Bacterial Gene Expression

The complexities of bacterial gene expression during mammalian infection cannot be addressed by in vitro experiments. We know that the infected host represents a complex and dynamic environment, which is modified during the infection process, presenting a variety of stimuli to which the pathogen must respond if it is to be successful. This response involves hundreds of ivi (in vivo– induced) genes which have recently been identified in animal and cell culture models using a variety of technologies including in vivo expression technology, differential fluorescence induction, subtractive hybridization and differential display. Proteomic analysis is beginning to be used to identify IVI proteins, and has benefited from the availability of genome sequences for increasing numbers of bacterial pathogens. The patterns of bacterial gene expression during infection remain to be investigated. Are ivi genes expressed in an organ–specific or cell–type–specific fashion ? New approaches are required to answer these questions. The uses of the immunologically based in vivo antigen technology system, in situ PCR and DNA microarray analysis are considered. This review considers existing methods for examining bacterial gene expression in vivo, and describes emerging approaches that should further our understanding in the future.

scholarly journals Transcription shifts in gut bacteria shared between mothers and their infants: data from the NiPPeR Study

2021 ◽  
Author(s):  
Tommi Vatanen ◽  
Olga Sakwinska ◽  
Brooke Wilson ◽  
Severine Combremont ◽  
Wayne Cutfield ◽  
...  
Keyword(s):  
Gene Expression ◽  
Large Scale ◽  
Gene Families ◽  
Gut Bacteria ◽  
Sequencing Data ◽  
Bacterial Strains ◽  
Bacterial Gene ◽  
Bacterial Gene Expression ◽  
Infant Gut Microbiome ◽  
Dependent Strain

Abstract The infant gut microbiome contains a portion of bacteria that originate from the maternal gut. In the infant gut these bacteria encounter a new metabolic environment that differs from the adult gut, consequently requiring adjustments in their activities. We utilized community RNA sequencing data (metatranscriptomes) from ten mother-infant dyads participating in the NiPPeR Study to characterize bacterial gene expression shifts following mother-to-infant transmission. Maternally derived bacterial strains adapted by large scale gene expression shifts following the transmission to the infant gut, with 12,564 activated and 14,844 deactivated gene families, including 1,007 transferases and 85 bacteriophage genes. The implicated genes were most numerous and the magnitude shifts greatest in Bacteroides spp. This study demonstrates environment-dependent, strain-specific shifts in gut bacteria function and underscores the importance of metatranscriptomic analysis in microbiome studies.

Impact of Genomic Technologies on Studies of Bacterial Gene Expression

2002 ◽  
Vol 56 (1) ◽  
pp. 599-624 ◽  
Author(s):  
Virgil Rhodius ◽  
Tina K. Van Dyk ◽  
Carol Gross ◽  
Robert A. LaRossa
Keyword(s):  
Gene Expression ◽  
Bacterial Gene ◽  
Bacterial Gene Expression ◽  
Genomic Technologies

Concurrence between the gene expression pattern of Actinobacillus actinomycetemcomitans in localized aggressive periodontitis and in human epithelial cells

2005 ◽  
Vol 54 (5) ◽  
pp. 497-504 ◽  
Author(s):  
Joseph Richardson ◽  
Justin Corey Craighead ◽  
Sam Linsen Cao ◽  
Martin Handfield
Keyword(s):  
Gene Expression ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Expression Pattern ◽  
Gene Expression Pattern ◽  
Aggressive Periodontitis ◽  
Actinobacillus Actinomycetemcomitans ◽  
Bacterial Gene ◽  
Bacterial Gene Expression

Actinobacillus actinomycetemcomitans is a facultatively intracellular pathogen and the aetiological agent of localized aggressive periodontitis. Screening of the genome of A. actinomycetemcomitans for in vivo-induced antigen determinants previously demonstrated that the proteome of this organism differs in laboratory culture compared with conditions found during active infection. The aim of the present study was to determine whether the bacterial gene expression pattern inferred with in vivo-induced antigen technology (IVIAT) in human infections was consistent with the gene expression pattern occurring upon epithelial cell association. To this end, a real-time PCR method was developed and used to quantify absolute and relative bacterial gene expression of A. actinomycetemcomitans grown extra- and intracellularly in two human epithelial cell lines (HeLa and IHGK). The amount of template used in the assay was normalized using the total count of viable bacteria (c.f.u.) as a reference point and performed in duplicate in at least two independent experiments. Controls for this experiment included 16S rRNA and gapdh. Transcription of all eight ORFs tested increased significantly (P < 0.05) in HeLa and IHGK cells compared with bacteria grown extracellularly. The concurrence of gene expression patterns found in the two models suggests that these epithelial cells are valid in vitro models of infection for the genes tested. IVIAT is an experimental platform that can be used as a validation tool to assess the reliability of animal and other models of infection and is applicable to most pathogens.

scholarly journals Regulation of asymmetries in the kinetics and protein numbers of bacterial gene expression

2019 ◽  
Vol 1862 (2) ◽  
pp. 119-128 ◽  
Author(s):  
Sofia Startceva ◽  
Vinodh K. Kandavalli ◽  
Ari Visa ◽  
Andre S. Ribeiro
Keyword(s):  
Gene Expression ◽  
Bacterial Gene ◽  
Bacterial Gene Expression

Metatranscriptomic analysis of lactic acid bacterial gene expression during kimchi fermentation

2013 ◽  
Vol 163 (2-3) ◽  
pp. 171-179 ◽  
Author(s):  
Ji Young Jung ◽  
Se Hee Lee ◽  
Hyun Mi Jin ◽  
Yoonsoo Hahn ◽  
Eugene L. Madsen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Lactic Acid ◽  
Bacterial Gene ◽  
Bacterial Gene Expression
Sign in / Sign up

Export Citation Format

Share Document