Sequence analysis of 23S rRNA gene and its amplification in bacteria discriminative identification

Objective: To study the difference of bacteria 23S rRNA gene and provide a tool for bacteria discriminative identification. Methods: The 23S rRNA gene fragment of different bacteria was amplified. The amplification fragments were identified with restriction enzymes analysis and sequencing. With bio-soft and the Gen-Bank data, the 23S rRNA conservative regions, mutant regions and their relation to the genetic development of bacterial strains were studied. Results: 23S rRNA gene fragment of 16 genera(species) bacteria was amplified and identified with restriction enzymes Hind III, Kpn I and SalI. The 23S rRNA gene fragment of 6 species were sequenced. 21 conservative regions were obtained. Conservative and mutant regions were separately distributed and most small mutant regions distributed like morsac among conservative regions. The phylogenetic trees among different bacteria drawn from the full lenth 23S rRNA gene or partial fragment were supported by results from 16S rRNA. Conclusion: The sequence distribution of 23S rRNA gene would be a solid base for the discriminative identification of bacteria strains and the construction of diagnosis gene chip.