Use of randomly amplified polymorphic DNA as a means of developing genus- and strain-specific Streptomyces DNA probes.
We have analyzed 20 randomly amplified polymorphic DNA (RAPD) primers against 36 Streptomyces strains, including 17 taxonomically undefined strains, 25 nonstreptomycete actinomycetes, and 12 outgroups consisting of gram-positive and -negative species. Most of the primers were useful in identifying unique DNA polymorphisms of all strains tested. We have used RAPD techniques to develop a genus-specific probe, one not necessarily targeting the ribosomal gene, for Streptomyces, and a strain-specific probe for the biological control agent Streptomyces lydicus WYEC108. In the course of these investigations, small-scale DNA isolations were also developed for efficiently isolating actinomycete DNA. Various modifications of isolation procedures for soil DNA were compared, and the reliability and specificity of the RAPD methodology were tested by specifically detecting the S. lydicus WYEC108 in DNA isolated from soil.
Roberts
M A
MA
Department of Microbiology, Molecular Biology, and Biochemistry, University of Idaho, Moscow, Idaho 83844-3052, USA.
Crawford
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DL
D015139:Blotting, Southern; D016172:DNA Fingerprinting; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique; D013045:Species Specificity; D013302:Streptomyces / Q000145:classification* / Q000235:genetics*
D015342:DNA Probes; D004269:DNA, Bacterial