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Gene function analysis in environmental isolates: The nif regulon of the strict iron oxidizing bacterium Leptospirillum ferrooxidans

AutorParro-García, Víctor ; Moreno-Paz, Mercedes
Palabras claveShotgun DNA microarrays
Gene expression
Nitrogen fixation
Iron oxidizer
Fecha de publicaciónjun-2003
EditorNational Academy of Sciences (U.S.)
CitaciónProc Natl Acad Sci U S A. 2003 June 24; 100(13): 7883–7888
ResumenA random genomic library from an environmental isolate of the Gram-negative bacterium Leptospirillum ferrooxidans has been printed on a microarray. Gene expression analysis was carried out with total RNA extracted from L. ferrooxidans cultures in the presence or absence of ammonium as nitrogen source under aerobic conditions. Although practically nothing is known about the genome sequence of this bacterium, this approach allowed us the selection and sequencing of only those clones bearing genes that showed an altered expression pattern. By sequence comparison, we have identified most of the genes of nitrogen fixation regulon in L. ferrooxidans, like the nifHDKENX operon, encoding the structural components of Mo-Fe nitrogenase; nifSU-hesB-hscBA-fdx operon, for Fe-S cluster assembly; the amtB gene (ammonium transporter); modA (molybdenum ABC type transporter); some regulatory genes like ntrC, nifA (the specific activator of nif genes); or two glnB-like genes (encoding the PII regulatory protein). Our results show that shotgun DNA microarrays are very powerful tools to accomplish gene expression studies with environmental bacteria whose genome sequence is still unknown, avoiding the time and effort necessary for whole genome sequencing projects.
DescripciónCopyright © by National Academy of Sciences. La versión original está disponible en http://www.pnas.org/
URIhttp://hdl.handle.net/10261/5056
DOI10.1073/pnas.1230487100
ISSN1091-6490
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