Nature of Polymorphisms in 16S-23S rRNA Gene Intergenic Transcribed Spacer Fingerprinting of Bacillus and Related Genera
Articolo
Data di Pubblicazione:
2003
Citazione:
Nature of Polymorphisms in 16S-23S rRNA Gene Intergenic
Transcribed Spacer Fingerprinting of Bacillus and
Related Genera / D. Daffonchio, A. Cherif, L. Brusetti, A. Rizzi, D. Mora, A. Boudabous, S. Borin. - In: APPLIED AND ENVIRONMENTAL MICROBIOLOGY. - ISSN 0099-2240. - 69:9(2003), pp. 5128-5137. [10.1128/AEM.69.9.5128-5137.2003]
Abstract:
The intergenic transcribed spacers (ITS) between the 16S and 23S rRNA genetic loci are frequently used in
PCR fingerprinting to discriminate bacterial strains at the species and intraspecies levels. We investigated the
molecular nature of polymorphisms in ITS-PCR fingerprinting of low-G C-content spore-forming bacteria
belonging to the genera Bacillus, Brevibacillus, Geobacillus, and Paenibacillus. We found that besides the
polymorphisms in the homoduplex fragments amplified by PCR, heteroduplex products formed during PCR
between amplicons from different ribosomal operons, with or without tRNA genes in the ITS, contribute to the
interstrain variability in ITS-PCR fingerprinting patterns obtained in polyacrylamide-based gel matrices. The
heteroduplex nature of the discriminating bands was demonstrated by fragment separation in denaturing
polyacrylamide gels, by capillary electrophoresis, and by cloning, sequencing, and recombination of purified
short and tRNA gene-containing long ITS. We also found that heteroduplex product formation is enhanced by
increasing the number of PCR cycles. Homoduplex-heteroduplex polymorphisms (HHP) in a conserved region,
such as the 16S and 23S rRNA gene ITS, allowed discrimination of closely related strains and species
undistinguishable by other methods, indicating that ITS-HHP analysis is an easy and reproducible additional
tool for strain typing.
Tipologia IRIS:
01 - Articolo su periodico
Elenco autori:
D. Daffonchio, A. Cherif, L. Brusetti, A. Rizzi, D. Mora, A. Boudabous, S. Borin
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