Nucleotide sequence and transcriptional analysis of the type a2 neurotoxin gene cluster in clostridium botulinum – documents

*Corresponding author. Clostridium botulinum signs and symptoms Tel.: +1-608-263-7944; fax: +1-608-263-

1 Present address: Department of Molecular Biology and Microbi-

The nucleotide sequences of the upstream regions of the botulinum neurotoxin type A1 (BoNT/A1) cluster of Clostridium

botulinum strain NCTC 2916 and the BoNT/A2 cluster of strain Kyoto-F were determined. Clostridium botulinum diagnosis A novel gene, designated orfx3, was

identi?ed following the orfx2 gene in both clusters. Clostridium botulinum intoxication ORF-X2 and ORF-X3 exhibit similarity to the BoNT cluster associated P-47

protein.


Clostridium botulinum culture media The BoNT/A1 and BoNT/A2 clusters share a similar gene arrangement, but exhibit di?erences in the spacing between

certain genes. Morphology of clostridium botulinum Sequences with similarity to transposases were identi?ed in these intergenic regions, suggesting that these di?erences

arose from an ancestral insertion event. Clostridium botulinum culture Transcriptional analysis of the BoNT/A2 cluster revealed that the genes of the cluster are

primarily synthesized as three polycistronic transcripts. Clostridium botulinum causes the disease known as botulism Two divergent polycistronic transcripts, one encoding the orfx1, orfx2, and

orfx3 genes, the second encoding the p47, ntnh, and bont/a2 genes, are transcribed from conserved BoNT cluster promoters. Clostridium botulinum structure The

third polycistronic transcript, expressed at low levels, encodes the positive regulatory botR gene and the orfx genes. Clostridium botulinum mode of transmission This is the ?rst

? 2004 Federation of European Microbiological Societies. Clostridium botulinum toxin type a Published by Elsevier B.V. What are the symptoms of clostridium botulinum All rights reserved.

Keywords: Clostridium botulinum; Botulinum neurotoxin; Toxin complex

ology, Tu?s University School of Medicine, 136 Harrison Avenue,

0378-1097/$22.00 ? 2004 Federation of European Microbiological Societies

Fig. Characteristics of clostridium botulinum 1. Clostridium botulinum life cycle Diagram of the BoNT/A clusters and ?anking regions in C. How is clostridium botulinum transmitted botulinu

Arrowheads show the binding position of primers (from Table 1) used for PC

aExperiment code: Kyoto-F sequencing (KS), NCTC 2916 sequencing (N

m strains. What is the significance of clostridium botulinum being anaerobic Open arrows represent the direction of gene transcription.

R. Clostridium botulinum temperature Primer pairs for PCR: 1/2, 3/4, 5/6, 8/36, 31/33, 32/34, 13/35. Pathogenesis of clostridium botulinum Lines

S.S. Definition of clostridium botulinum Dineen et al. How do you get clostridium botulinum / FEMS Microbiology Letters 235 (2004) 9–16 13

Fig. Clostridium botulinum cdc 2. Clostridium botulinum aerobic or anaerobic Northern blot analyses of BoNT/A2 cluster gene expression. What does clostridium botulinum need to thrive T

electrophoresis, transferred to nylon membranes, and hybridized with g

of RNA molecular size markers is shown ?anking the ?gure. Causado pela bacteria clostridium botulinum Arrowhead

representing total RNA isolated from two independent cultures.

NA isolated from C. Clostridium botulinum symptoms and signs botulinum strain Kyoto-F was separated by gel

eci?c 32P-radiolabeled probes (listed above each panel). What disease is caused by clostridium botulinum The migration

w the positions of the major transcripts. Caracteristicas de la bacteria clostridium botulinum Each panel contains two lanes

14 S.S. Clostridium botulinum bacteria information Dineen et al. La bacteria clostridium botulinum / FEMS Microbiology Letters 235 (2004) 9–16

Fig. Clostridium botulinum symptoms food poisoning 3. Clostridium botulinum intoxication symptoms Comparison of neurotoxin gene cluster putative promoter regi

BoNT/A2, BoNT/A1 (strain NCTC 2916), BoNT/B [13], and TeNT [2

precedes is shown in the ?rst column. Clostridium botulinum c2 toxin Where data are available, transcr

represent conserved sequences in the promoter regions shown. What is clostridium botulinum bacteria The co

putative promoters preceding the botR gene in BoNT/A2 and BoNT/B [13]

(A) Alignment of the putative promoters for the major transcripts of

ne clusters. D value clostridium botulinum The gene and cluster for which each nucleotide sequence

initiation sites are indicated by ‘‘blocked’’ nucleotides. What does clostridium botulinum eat Boxes 1 and 2

us (CONS) sequence is shown in the last row. What does clostridium botulinum look like (B) Alignment of the

clusters. What does clostridium botulinum mean Potential )10 and )35 promoter elements are indicated.

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16 S.S. Definition of clostridium botulinum Dineen et al. How do you get clostridium botulinum / FEMS Microbiology Letters 235 (2004) 9–16

Nucleotide sequence and transcriptional analysis of the type A2 neurotoxin gene cluster in Clostridium botulinum

Sequence analysis of the BoNT/A2 cluster of strain Kyoto-F and the BoNT/A1 cluster of strain NCTC 2916

Transcriptional analysis of the BoNT/A2 cluster in strain Kyoto-F

Determination of the transcription start points and promoters of the BoNT/A2 cluster genes

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