Use of RNA-arbitrarily-primed PCR to detect the induction of gene expression in freshwater bivalves (Unio tumidus) transplanted into the Moselle River

2004 ◽  
Vol 43 (4) ◽  
pp. 209-216 ◽  
Author(s):  
Fran�ois Rodius ◽  
Claude Hammer ◽  
Paule Vasseur
Keyword(s):  
Gene Expression ◽  
Unio Tumidus ◽  
Freshwater Bivalves ◽  
Arbitrarily Primed Pcr

scholarly journals Identification of Differential Gene Expression in Bacteria Associated with Coral Black Band Disease by Using RNA-Arbitrarily Primed PCR

2004 ◽  
Vol 70 (6) ◽  
pp. 3687-3694 ◽  
Author(s):  
Jorge Frias-Lopez ◽  
George T. Bonheyo ◽  
Bruce W. Fouke
Keyword(s):  
Gene Expression ◽  
Differential Gene Expression ◽  
Black Band Disease ◽  
Expression Of Genes ◽  
Black Band ◽  
Differential Gene ◽  
Apoprotein A1 ◽  
Arbitrarily Primed Pcr ◽  
Transporter Systems ◽  
First Time

ABSTRACT RNA-arbitrarily primed PCR techniques have been applied for the first time to identify differential gene expression in black band disease (BBD), a virulent coral infection that affects reef ecosystems worldwide. The gene activity for the BBD mat on infected surfaces of the brain coral Diploria strigosa was compared with that for portions of the BBD mat that were removed from the coral and suspended nearby in the seawater column. The results obtained indicate that three genes (DD 95-2, DD 95-4, and DD 99-9) were up-regulated in the BBD bacterial mat on the coral surface compared to the transcript base levels observed in the BBD mat suspended in seawater. Clone DD 95-4 has homology with known amino acid ABC transporter systems in bacteria, while clone DD 99-9 exhibits homology with chlorophyll A apoprotein A1 in cyanobacteria. This protein is essential in the final conformation of photosystem I P700. DD 95-2, the only gene that was fully repressed in the BBD mat samples suspended in seawater, exhibited homology with the AraC-type DNA binding domain-containing proteins. These transcriptional activators coordinate the expression of genes essential for virulence in many species of gram-negative bacteria.

scholarly journals Identification of Aerobically and Anaerobically Induced Genes in Enterococcus faecalis by Random Arbitrarily Primed PCR

1999 ◽  
Vol 65 (4) ◽  
pp. 1470-1476 ◽  
Author(s):  
Brett D. Shepard ◽  
Michael S. Gilmore
Keyword(s):  
Gene Expression ◽  
Enterococcus Faecalis ◽  
Oxygen Metabolism ◽  
Differentially Expressed ◽  
Bacterial Rna ◽  
Eukaryotic Gene ◽  
Differential Gene ◽  
Pcr Method ◽  
Induced Genes ◽  
Arbitrarily Primed Pcr

ABSTRACT Enterococci have emerged among the leading causes of nosocomial infection. With the goal of analyzing enterococcal genes differentially expressed in environments related to commensal or environmental colonization and infection sites, we adapted and optimized a method more commonly used in the study of eukaryotic gene expression, random arbitrarily primed PCR (RAP-PCR). The RAP-PCR method was systematically optimized, allowing the technique to be used in a highly reproducible manner with gram-positive bacterial RNA. In the present study, aerobiosis was chosen as a variable for the induction of changes in gene expression by Enterococcus faecalis. Aerobically and anaerobically induced genes were detected and identified to the sequence level, and differential gene expression was confirmed by quantitative, specifically primed RT-PCR. Differentially expressed genes included several sharing identity with those of other organisms related to oxygen metabolism, as well as hypothetical genes lacking identity to known genes.

scholarly journals Arbitrarily primed PCR fingerprinting of RNA and DNA in Entamoeba histolytica

2000 ◽  
Vol 42 (5) ◽  
pp. 249-253 ◽  
Author(s):  
Paulo Renato VALLE ◽  
Maria Betânia G. SOUZA ◽  
Edna M. PIRES ◽  
Edward F. SILVA ◽  
Maria A. GOMES
Keyword(s):  
Gene Expression ◽  
Rapd Analysis ◽  
Polymorphic Markers ◽  
Biological Features ◽  
Pcr Products ◽  
The Individual ◽  
Arbitrarily Primed Pcr ◽  
High Degree ◽  
Rna And Dna

Differences were detected in the gene expression of strains of E. histolytica using RNA (RAP-PCR) and DNA fingerprinting (RAPD). Analysis of the electrophoretic profiles of the gels revealed some polymorphic markers that could be used in the individual characterization of the strains. The 260 bands generated by using five different primers for RAP-PCR, as well as RAPD, were employed in the construction of dendograms. The dendogram obtained based on the RAPD products permitted the distinction of symptomatic and asymptomatic isolates, as well the correlation between the polymorphism exhibited and the virulence of the strains. The dendogram obtained for the RAP-PCR products did not show a correlation with the virulence of the strains but revealed a high degree of intraspecific transcriptional variability that could be related to other biological features, whether or not these are involved in the pathogenesis of amebiasis.

scholarly journals Human waterborne protozoan parasites in freshwater bivalves (Anodonta anatina and Unio tumidus) as potential indicators of fecal pollution in urban reservoir

Limnologica ◽  
2015 ◽  
Vol 51 ◽  
pp. 32-36 ◽  
Author(s):  
Anna Słodkowicz-Kowalska ◽  
Anna C. Majewska ◽  
Piotr Rzymski ◽  
Łukasz Skrzypczak ◽  
Anna Werner
Keyword(s):  
Fecal Pollution ◽  
Protozoan Parasites ◽  
Unio Tumidus ◽  
Freshwater Bivalves

scholarly journals RNA Arbitrarily Primed PCR Survey of Genes Regulated by ToxR in the Deep-Sea Bacterium Photobacterium profundum Strain SS9

2001 ◽  
Vol 183 (5) ◽  
pp. 1688-1693 ◽  
Author(s):  
Kelly A. Bidle ◽  
Douglas H. Bartlett
Keyword(s):  
Gene Expression ◽  
Virulence Gene ◽  
Environmental Signals ◽  
Virulence Gene Expression ◽  
New Genes ◽  
Protein Encoding ◽  
Arbitrarily Primed Pcr ◽  
Photobacterium Profundum ◽  
Encoding Genes

ABSTRACT We are currently investigating the role of ToxR-mediated gene regulation in Photobacterium profundum strain SS9. SS9 is a moderately piezophilic (“pressure loving”) psychrotolerant marine bacterium belonging to the family Vibrionaceae. InVibrio cholerae, ToxR is a transmembrane DNA binding protein involved in mediating virulence gene expression in response to various environmental signals. A homolog to V. choleraeToxR that is necessary for pressure-responsive gene expression of two outer membrane protein-encoding genes was previously found in SS9. To search for additional genes regulated by ToxR in SS9, we have used RNA arbitrarily primed PCR (RAP-PCR) with wild-type and toxRmutant strains of SS9. Seven ToxR-activated transcripts and one ToxR-repressed transcript were identified in this analysis. The cDNAs corresponding to these partial transcripts were cloned and sequenced, and ToxR regulation of their genes was verified. The products of these genes are all predicted to fall into one or both of two functional categories, those whose products alter membrane structure and/or those that are part of a starvation response. The transcript levels of all eight newly identified genes were also characterized as a function of hydrostatic pressure. Various patterns of pressure regulation were observed, indicating that ToxR activation or repression cannot be used to predict the influence of pressure on gene expression in SS9. These results provide further information on the nature of the ToxR regulon in SS9 and indicate that RAP-PCR is a useful approach for the discovery of new genes under the control of global regulatory transcription factors.

scholarly journals Proteomic Analysis of Interactions between a Deep-Sea Thermophilic Bacteriophage and Its Host at High Temperature

2009 ◽  
Vol 84 (5) ◽  
pp. 2365-2373 ◽  
Author(s):  
Dahai Wei ◽  
Xiaobo Zhang
Keyword(s):  
Gene Expression ◽  
High Temperature ◽  
Virus Infection ◽  
Differentially Expressed Genes ◽  
Deep Sea ◽  
Proteomic Analysis ◽  
Differentially Expressed ◽  
Host Responses ◽  
Host Interaction ◽  
Arbitrarily Primed Pcr

ABSTRACT The virus-host interaction is essential to understanding the role that viruses play in ecological and geochemical processes in deep-sea vent ecosystems. Virus-induced changes in cellular gene expression and host physiology have been studied extensively. However, the molecular mechanism of interaction between a bacteriophage and its host at high temperature remains poorly understood. In the present study, the virus-induced gene expression profile of Geobacillus sp. E263, a thermophile isolated from a deep-sea hydrothermal ecosystem, was characterized. Based on proteomic analysis and random arbitrarily primed PCR (RAP-PCR) of Geobacillus sp. E263 cultured under non-bacteriophage GVE2 infection and GVE2 infection conditions, there were two types of protein/gene profiles in response to GVE2 infection. Twenty differentially expressed genes and proteins were revealed that could be grouped into 3 different categories based on cellular function, suggesting a coordinated response to infection. These differentially expressed genes and proteins were further confirmed by Northern blot analysis. To characterize the host proteins in response to virus infection, aspartate aminotransferase (AST) was inactivated to construct the AST mutant of Geobacillus sp. E263. The results showed that the AST protein was essential in virus infection. Thus, transcriptional and proteomic analyses and functional analysis revealed previously unknown host responses to deep-sea thermophilic virus infection.

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