Rapid characterization by colony direct PCR of distribution specificity in Streptomyces of kan gene encoding a specific aminoglycoside-3-N-acetyltransferase.

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(Using colony direct PCR we established, kan gene (GenBank accession No. AB028210) coding for an aminoglycoside acetyltransferase, AAC(3), was surveyed among strains of Streptomyces griseus and its related taxa (S. anulatus group). The regulatory and ORF regions of kan gene were targeted by the use of original primers. With the regulatory region primers, the target fragment (518 bp) amplification was observed in all the streptomycin (SM)-producing strains of S. griseus and in several strains of S. anulatus group. The other target fragment (930 bp) amplification using the ORF primers was observed only in the SM-producing strains of S. griseus including ‘S. ornatus’ that has substantially identical properties to those of S. griseus. It was thus conclusive that kan gene is highly conserved in SM-producing strains of S. griseus and distributed in taxa related to S. griseus. There were numbers of strains that provided no amplification upon PCR but hybridization bands upon Southern hybridization, suggesting the lack or alteration of kan gene. Characterization by colony direct PCR using appropriate primers will be a big help for taxonomic as well as ecological characterization of actinomycetes.

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