Open AccessUse of Randomly Amplified Polymorphic DNA as a Means of Developing Genus- and Strain-Specific Streptomyces DNA Probes
Author(s) -
Mark A. Roberts,
Don L. Crawford
Publication year - 2000
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.66.6.2555-2564.2000
Subject(s) - rapd , biology , dna , streptomyces , genetics , strain (injury) , dna sequencing , hybridization probe , streptomycetaceae , actinomycetales , microbiology and biotechnology , bacteria , population , demography , anatomy , sociology , genetic diversity
We have analyzed 20 randomly amplified polymorphic DNA (RAPD) primers against 36Streptomyces 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, forStreptomyces , and a strain-specific probe for the biological control agentStreptomyces 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 theS. lydicus WYEC108 in DNA isolated from soil.