Heterogeneity in Bacillus cereus PCR products detected by ESI-FTICR mass spectrometry

Analytical Chemistry
D S WunschelR D Smith

Abstract

PCR amplification of a segment of the 16/23S rDNA interspace region (ISR) from Bacillus cereus 6464 produced a mixture of products. An 89-bp product was predicted on the basis of the reported sequence. The ESI-FTICR analysis revealed three double-stranded products, differing in size by a single nucleotide corresponding to two homoduplexes of 89 and 88 base pairs and a heteroduplex of 89 and 88 nucleotide strands. These were produced from a single preparation of genomic DNA and a single primer pair. ESI-FTICR analysis of the single strands identified a deletion of a T in the coding strand and a corresponding loss of an A in the noncoding strand of this product. The ESI-FTICR analysis indicated the presence of an unreported sequence variation between rRNA operons in this organism. This report illustrates that PCR products amplified from templates differing by a single nucleotide can be resolved and identified using ESI-FTICR at the 89-bp level. Furthermore, the ESI-FTICR mass measurements provided the identity of the deletion, which is indicative of interoperon variability.

References

Mar 11, 1982·Nucleic Acids Research·K LoughneyJ E Dahlberg
Jul 1, 1995·Journal of Clinical Microbiology·L J HarrellK H Wilson
Jan 1, 1997·Rapid Communications in Mass Spectrometry : RCM·T TsuneyoshiD J Prockop

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Citations

Apr 1, 2003·Mass Spectrometry Reviews·Jörg Tost, Ivo G Gut
Jan 13, 2004·Biomolecular Engineering·Zhaojing MengPatrick A Limbach
Jan 5, 2002·Journal of the American Society for Mass Spectrometry·Allison P Null, David C Muddima
Nov 30, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·James J WaltersA Fox
Sep 27, 2002·Applied and Environmental Microbiology·Gert B JensenLars Andrup
Dec 16, 2003·Rapid Communications in Mass Spectrometry : RCM·Allison P NullDavid C Muddiman
Sep 25, 2004·Mass Spectrometry Reviews·Steven A HofstadlerJames C Hannis
Jul 9, 2004·Chembiochem : a European Journal of Chemical Biology·David S WunschelRichard D Smith
Apr 11, 2000·Journal of the American Society for Mass Spectrometry·D S WunschelR D Smith
Apr 16, 2002·Journal of the American Society for Mass Spectrometry·Allison P NullDavid C Muddiman

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