Abstract
With the developments in taxonomy, the classically used highly conserved 16S rRNA molecular marker has shown some disadvantages among closely related species. For further taxonomic studies of the prokaryotes, specific PCR primers were designed from two conserved regions in the amino acid sequences of the 70-kDa heat shock protein sourced from 20 different genera in actinomycetes. These were used for the amplification of the hsp70 genes in 16 Streptomyces strains. Then, we investigated the phylogenetic relationships among these Streptomyces strains and compared the tree topology based on the hsp70 gene with those based on the previously used markers (16S rRNA and gyrB). To our knowledge, this is the first use of the hsp70 gene as a molecular marker for the taxonomic identification of Streptomyces.
Highlights
The highly conserved 16S rRNA gene, known as the ‘bacterial fossil’, encodes small subunit of the ribosomal RNA in prokaryotes
Design primers for the polymerase chain reaction (PCR) amplification of hsp70 genes PCR primers for the amplification of hsp70 (Dna K) genes were designed from two conserved regions of the amino acid sequences of the 70-kDa heat shock protein from 20 genera in the actinomycetes (Fig. 1)
The results showed that the sequences of the PCR product amplified from the primer pair was the same as the hsp70 genes retrieved from the Genbank database
Summary
The highly conserved 16S rRNA gene, known as the ‘bacterial fossil’, encodes small subunit of the ribosomal RNA (rRNA) in prokaryotes. It contains variable regions and constant regions (Woese et al 1975). Phylogenetic analysis based on the 16S rRNA gene is considered to be a simple and appropriate tool for the construction of bacterial phylogenetic relationships. It is one of the most commonly used methods for identifying microorganisms (Amann et al 1995; Stackebrandt and Goebel 1994; Woese et al 1990). The high percentage of sequence similarity among closely related species limits its effectiveness (Ash et al 1991; Christensen et al 1998; Yamamoto and Harayama 1998)
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