A prototype taxonomic microarray targeting the rpsA housekeeping gene permits species identification within the rhizobial genus Ensifer.
To develop a reliable tool for the identification and classification of the different Ensifer species, without the need for sequencing, a prototype DNA microarray that targets the rpsA housekeeping gene was designed and tested. Internal segments of the rpsA gene from 34 reference strains, representing the different Ensifer species, were sequenced and the sequences were used to select 44 diagnostic oligonucleotides that served as probes for the identification microarray. Both, genomic DNA and specific rpsA PCR-products were tested as a target in hybridisation experiments. Experimental conditions were optimised and the diagnostic oligonucleotides were validated. Hybridisation results with the rpsA PCR-products showed reliable identification of the reference strains to species and genomovar level. Our data indicate that a microarray targeting housekeeping genes is a promising, accurate and relatively simple genotyping technique that would also be applicable for the identification and characterization of other bacterial groups of interest.
Martens
Miet
M
Laboratorium voor Microbiologie (WE10), Universiteit Gent, B-9000 Ghent, Belgium.
Weidner
Stefan
S
Linke
Burkhard
B
de Vos
Paul
P
Gillis
Monique
M
Willems
Anne
A
D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000032:analysis* / Q000235:genetics; D012230:Rhizobiaceae / Q000145:classification* / Q000235:genetics; D012269:Ribosomal Proteins / Q000235:genetics* / Q000378:metabolism
D012336:RNA, Ribosomal, 16S; D012269:Ribosomal Proteins; C015494:ribosomal protein S1