Identification of aerobically and anaerobically induced genes in Enterococcus faecalis by random arbitrarily primed PCR.
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.
Shepard
B D
BD
Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA.
Gilmore
M S
MS
D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism; D000332:Aerobiosis; D000693:Anaerobiosis; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D013293:Enterococcus faecalis / Q000235:genetics* / Q000254:growth & development* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology*; D006801:Humans; D009245:NADH Dehydrogenase / Q000235:genetics / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D012329:RNA, Bacterial / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012714:Serine-tRNA Ligase / Q000235:genetics / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; D012329:RNA, Bacterial; D010088:Oxidoreductases; D002374:Catalase; D009245:NADH Dehydrogenase; D012714:Serine-tRNA Ligase