• Journal of Microbiology
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• v.33 no.2
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• pp.95-102
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• 1995

A probability identification matrix of acidophilic Streptomyces was constructed. The phenetic data of the strains were derived from numerical classification described by Seong et al. The minimum number of diagnostic characters was determined using computer programs for calculation of different separation indices. The resulting matrix consisted of 25 clusters versus 53 characters. Theoretical evaluation of this matrix was achieved by estimating the chuster overlap and the identification scores for the Hypothetical Median Organisms (HMO) and for the representatives of each cluster. Cluster overlap was found to be relatively small. Identification scores for the HMO and the randomly selected representatives of each cluster were satisfactory. The matrix was assessed practically by applying the matrix to the identification of unknown isolates. Of the unknown isolates, 71.9% were clearly identified to one of eight clusters. The numerical classification data was also used to design a selective isolation medium for antibiotic-producing organisms. Four chemical substances including 2 antibiotics were determined by the DLACHAR program as diagnostic for the isolation of target organisms which have antimicrobial activity against Micrococcus luteus. It was possible to detect the increased rate of selective isolation on the synthesized medium. Theresults show that the numerical phenetic data can be applied to a variety of purposes, such as construction of identification matrix and selective isolation medium for acidophilic antinomycetes.

• Journal of Microbiology
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• v.33 no.2
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• pp.142-145
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• 1995

The pumb gene encoding a basic amino acid transport protein in Neurospora crassa could be cloned by using a mutant strain defective in pmb gene as a host strain, using a negative selection on the media containing amino acid analogue canavanine. To select positive transformants of the genes for cloning, an auxotrophic marker (his-2) was added to a pmb mutant strain by mating ; a triple mutant (pmn : pmb : his-2) was constructued by crossing a strain defective in basic amino acid transport system (# 1683-bat um 535 "A") to a double mutant strain defective in neutral amino acid transport and histidine production (mitrol : his-2 "a"). Crossing was performed on synthetic crossing (SC) media containing histidine. The pmn : pmb and pmn :pmb : his-2 strains were selected among the progeny colonies from crosses on plates containing 5- .mu.g/ml para-fluoro-phenylalanine (PFPA), 200 .mu.g/ml canavanine, and 500 .mu.g/ml histidine. The selected colonies were cultured on minimal media with or without histidine for discarding pmn : pmb strain, because the pmn : pmb : his -2 strain grows only on histidine containing media. The pmn :pmb : his-2 strain selected can be used as a host strain for the cloning of the pmb and the pmn genes from a Neurospora genomic library by means of positive selections.