Molecular characterization of thermophilic Streptomyces using amplified ۱۶s ribosomal DNA restriction analysis (ARDRA)

The present study aims to characterize the ۱۹ thermophilic Streptomyces isolates on the basis of their restriction profile obtained by Amplified Ribosomal DNA Restriction Analysis (ARDRA) method. Nineteen thermophilic Streptomyces isolates, characterized and identified on the basis of cultural, morphological, physiological and biochemical characteristics using PIB Win (Probabilistic identification of Bacteria) software, have been further characterized by Amplified Ribosomal DNA Restriction Analysis method. DNA of all the ۱۹ selected isolates and two reference Streptomyces cultures of MTCC (S. phaeochromogenes (MTCC ۲۶۱۴) and S. thermovulgaris (MTCC ۱۸۲۲) were isolated and the ۱۶S rRNA gene was amplified using primer F۱ (۵'-AGAGTTTGATCITGGCTCAG-۳’; I= inosine) and primer R۵ (۵'-ACGGITACCTTGTTACGACTT-۳'). Two restriction endonuclease enzymes i.e. Eco R I and Hae III were used for restriction digestion. The ۱۶S rDNA were amplified by PCR using specific primers and the amplified product were cleaved separately by both endonucleases, yielding ۴ to ۵ bands by Hae III and ۲ to ۳ bands by Eco R I. Digestion with two endonucleases indicate that Hae III enzyme is better than Eco R I for differentiating Streptomyces isolates.