The possibility of discriminating within the Bacillus cereus group using gyrB sequencing and PCR-RFLP.
Based on a combination of PCR and restriction endonuclease (RE) digestion (PCR-RE digestion), we have examined the possibility of differentiating members of the Bacillus cereus group. Fragments of the gyrB gene (362 bp) from pure cultures of 12 B. cereus, 25 B. thuringiensis, 25 B. mycoides and two B. anthracis strains were amplified and subsequently digested with Sau3A1. Furthermore, a majority of the amplicons were sequenced directly to verify the PCR-RE results. The results obtained suggest that only the B. mycoides generates specific fragments following PCR-RE. Conversely, it was not possible to discriminate between the B. cereus and the B. thuringiensis strains using the methods described.
Jensen
Gert B
GB
Thomsen Bioscience A/S, Strømmen 6, DK-9400 Nørresundby, Denmark. gbj@thomsen-bioscience.com
Fisker
Niels
N
Sparsø
Thomas
T
Andrup
Lars
L
D001409:Bacillus cereus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D027081:DNA Gyrase / Q000235:genetics*; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000032:analysis*; D005516:Food Microbiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D015183:Restriction Mapping; D013045:Species Specificity
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes; D027081:DNA Gyrase