Identification of some actinomycete species by restrication endonuclease analysis of an amplified the 16SrRNA gene

Abstract

Restriction fragment length polymorphisms of 16S rRNA genes of representative strains of the actinomycete species (P-100-1 Saccharomonospora halophila, A-3-4 Nocardiopsis sp., A-1 Nocardiopsis halotolerance, P-7-2 Nocradiopsis sp. and Microbispora sp.) were determined following the isolation and genes amplification. 16S rDNAs were amplified with two primers; 5\'-TCACGGAGAGTTTGATCCTG-3\' and 5\'-AGAAAGGAGGTGATC-3\'. Actinomycete 16S rRNA genes were used with eight restriction endonucleases (EcoRI, StyI, AvaI, BanI, HindIII, NaeI, AsnI and BanII) to obtain restriction fragment length polymorphisms (RFLPs). The PCR/RFLP profiles of SrRNA genes indicated that the actinomycete isolates are polyphylogenetic when digested with BanI restriction endonuclease. Also, some actinomycete strains are monophylogenetic in nature, such as N. A-3-4 and S. P-100-1. PCR-RFLPs of SrDNA is an effective marker to differentiate between the actinomycete strains in a much more accuracy than do the morphological studies alone.

Egyptian Journal of Biotechnology Vol. 25 (1) 2007: pp. 13-26