Microarray-based detection and typing of the Rhizobium nodulation gene nodC: potential of DNA arrays to diagnose biological functions of interest.
Environmental screening of bacteria for the presence of genes of interest is a challenging problem, due to the high variability of the nucleotide sequence of a given gene between species. Here, we tackle this general issue using a particularly well-suited model system that consists of the nodulation gene nodC, which is shared by phylogenetically distant rhizobia. 41mer and 50mer oligonucleotides featuring the nucleotide diversity of two highly conserved regions of the NodC protein were spotted on glass slides and cross hybridized with the radioactive-labeled target genomic DNA under low-stringency conditions. Statistical analysis of the hybridization patterns allowed the detection of known, as well as new, nodC sequences and classified the rhizobial strains accordingly. The microarray was successfully used to type the nodC gene directly from legume nodules, thus eliminating the need of cultivation of the endosymbiont. This approach could be extended to a panel of diagnostic genes and constitute a powerful tool for studying the distribution of genes of interest in the environment, as well as for bacteria identification.
Bontemps
Cyril
C
Laboratoire des Interactions Plantes Micro-Organismes, INRA-CNRS, BP 52627, 31326 Castanet-Tolosan cedex, France.
Golfier
Geoffroy
G
Gris-Liebe
Carine
C
Carrere
Sébastien
S
Talini
Luc
L
Boivin-Masson
Catherine
C
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D015373:Bacterial Typing Techniques; D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D005810:Multigene Family; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D010802:Phylogeny; D012231:Rhizobium / Q000145:classification / Q000201:enzymology / Q000235:genetics*
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D017351:N-Acetylglucosaminyltransferases; C060771:NodC protein, Rhizobiales