Cloning and characterization of the upstream region of Clostridium botulinum type B neurotoxin gene

Yang, Gi-Hyeok; Rhee, Sang Dal; Jung, Hyun Ho; Jhee, Ok Hwa; Yang, Kyu Hwan

doi:10.1080/15216549800202782

Cited by 2 publications

(4 citation statements)

References 3 publications

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“…Northern analyses of RNA isolated from these strains (Fig. 3) showed two type B-speci¢c transcripts with sizes (4 and 6 kb) corresponding approximately to those reported previously for the type A gene [16], and slightly smaller than those reported for the type B gene [17]. Several poorly resolved bands ( 9 1.9 kb) were also detected and attributed to degradation.…”

Section: Expression Analyses Of the Type B Neurotoxin Mrnasupporting

confidence: 70%

“…To determine whether this might be the case, we compared the levels of type B toxin-speci¢c mRNA recovered from strains 588 and 593. Thus, di¡erences responsible for the phenotypic di¡erences in expression must be localized to a more distal region of the genome, possibly from a transcriptional readthrough from a second more distal promoter that directs expression of the NTNH gene located immediately upstream of the neurotoxin gene [17]. 3) showed two type B-speci¢c transcripts with sizes (4 and 6 kb) corresponding approximately to those reported previously for the type A gene [16], and slightly smaller than those reported for the type B gene [17].…”

Section: Expression Analyses Of the Type B Neurotoxin Mrna For Strainmentioning

confidence: 99%

“…4). Thus, di¡erences responsible for the phenotypic di¡erences in expression must be localized to a more distal region of the genome, possibly from a transcriptional readthrough from a second more distal promoter that directs expression of the NTNH gene located immediately upstream of the neurotoxin gene [17].…”

Section: Expression Analyses Of the Type B Neurotoxin Mrnamentioning

confidence: 99%

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Characterization of six type A strains ofClostridium botulinumthat contain type B toxin gene sequences

Kirma

Ferreira

Baumstark

2004

FEMS Microbiology Letters

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Six Clostridium botulinum isolates exhibiting type A toxicity as measured by the mouse bioassay were found to contain both type A and type B neurotoxin DNA sequences. The six strains were divided into three groups based on the DNA sequence of the type B neurotoxin gene. Members of each group exhibited 100% sequence identity over the 3876 bp type B toxin open reading frame. The type B toxin sequence of all groups differed at more than 60 positions when compared to the BGB control strain.

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Section: Expression Analyses Of the Type B Neurotoxin Mrnasupporting

confidence: 70%

Section: Expression Analyses Of the Type B Neurotoxin Mrna For Strainmentioning

confidence: 99%

Section: Expression Analyses Of the Type B Neurotoxin Mrnamentioning

confidence: 99%

See 1 more Smart Citation

Characterization of six type A strains ofClostridium botulinumthat contain type B toxin gene sequences

Kirma

Ferreira

Baumstark

2004

FEMS Microbiology Letters

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“…3, lane 2) were similar to those of 12S and 16S toxins, respectively, of other serotypes (2,4,6,12,13), and the N-terminal sequences were identical to those deduced from the nucleotide sequences of the serotype B toxin genes previously published (Fig. 3) (15,19,20,21). Thus, it was concluded that the molecular compositions of serotype B 12S and 16S toxins are similar to those of other serotypes.…”

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confidence: 56%

Purification of Fully ActivatedClostridium botulinumSerotype B Toxin for Treatment of Patients with Dystonia

et al. 2003

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Clostridium botulinum serotype B toxins 12S and 16S were separated by using a beta-lactose gel column at pH 6.0; toxin 12S passed through the column, whereas toxin 16S bound to the column and eluted with lactose. The fully activated neurotoxin was obtained by applying the trypsin-treated 16S toxin on the same column at pH 8.0; the neurotoxin passed through the column, whereas remaining nontoxic components bound to the column. The toxicity of this purified fully activated neurotoxin was retained for a long period by addition of albumin in the preparation.Clostridium botulinum strains produce immunologically distinct neurotoxins (serotypes A to G). The molecular masses of neurotoxin types A to G are approximately 150 kDa. The neurotoxins are produced as a single form and become dichain-form light (50-kDa) and heavy (100-kDa) chains by cleavage with proteases such as trypsin at about one-third of the distance from the amino terminus, and the toxic activity of the dichain form becomes fully activated (1). In culture fluid and food with acidic conditions, the neurotoxins associate with nontoxic components and form large complexes designated progenitor toxins. Under alkaline conditions, the progenitor toxins dissociate into neurotoxin and nontoxic components (10, 18). The progenitor toxins are found in three forms with molecular masses of 900 kDa (19S), 500 kDa (16S), and 300 kDa (12S) (14). The 12S toxin is composed of a neurotoxin and a nontoxic component having no hemagglutinin (HA) activity (designated nontoxic non-HA [NTNH]), whereas the 16S and 19S toxins are composed of a neurotoxin, NTNH, and HA. The serotype A strain produces three forms of toxins (19S, 16S, and 12S). Type B, C, and D strains produce the 16S and the 12S toxins. We purified different-sized progenitor toxins from serotype A, C, and D cultures (2,4,6,12,13) and demonstrated that (i) the 19S toxin is a dimer of the 16S toxin; (ii) HA consists of four subcomponents with molecular masses of 52 to 53, 33 to 35, 19 to 23, and 15 to 17 kDa, designated here as HA3b, HA1, HA3a, and HA2, respectively; and (iii) NTNH of the 12S toxin has a cleavage site(s) at the N-terminal region.Recently, serotype A and B progenitor toxins have been used for treating patients with strabismus, blepharospasm, nystagmus, facial spasm, spastic aphonia, and many other forms of dystonia (9, 11). In both toxin types, progenitor toxins are used because they are easily obtained and are more stable than neurotoxin. The treatment is very effective but has a serious side effect for some patients in whom antiprogenitor toxins, including antineurotoxin antibodies, are produced after several injections. It seems that using neurotoxin alone is better than using the progenitor toxin (a complex of the neurotoxin and a nontoxic component). Furthermore, it has been reported that serotype B toxin, which is used therapeutically at present, is partially cleaved (16) and therefore the toxin is not fully activated. In this paper, we report a simple procedure for largescale purification of botu...

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Cloning and characterization of the upstream region of Clostridium botulinum type B neurotoxin gene

Cited by 2 publications

References 3 publications

Characterization of six type A strains ofClostridium botulinumthat contain type B toxin gene sequences

Characterization of six type A strains ofClostridium botulinumthat contain type B toxin gene sequences

Purification of Fully ActivatedClostridium botulinumSerotype B Toxin for Treatment of Patients with Dystonia

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