Cloning and expression of the dermonecrotic toxin gene of Pasteurella multocida ssp. multocida in Echerichia coli

Kamps, A. M. I. E.

doi:10.1016/0378-1097(90)90192-s

Cited by 4 publications

(5 citation statements)

References 4 publications

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“…Only total sequencing of each toxin or toxin gene in a phylogenetic examination could reveal possible minor differences. The PMT-encoding gene from two type D strains of P. multocida have now been sequenced and the deduced amino acid sequences diverge only in 3 amino acid residues (12,18). This provides further evidence that toxin from type A strains and type D strains are serologically and antigenetically closely related, if not identical.…”

Section: Discussionmentioning

confidence: 79%

Characterization of Toxin from Different Strains of Pasteurella multocida Serotype A and D

Frandsen

Foged

Petersen

et al. 1991

Journal of Veterinary Medicine, Series B

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Summary Chromosomal DNA from 13 different selected Pasteurella multocida spp. multocida strains of serotypes A and D were isolated and compared. All 10 toxigenic strains were recognized by a DNA probe which included the toxA gene coding for the Pasteurella multocida toxin (PMT). None of the three nontoxigenic strains reacted with the DNA probe. Toxin from the 10 toxigenic strains were isolated and compared. All were found to possess the biological characteristics previously described for the PMT isolated from P. multocida ssp. multocida NCTC 12178, including molecular mass of approx. 143 kDa and reactivity with a series of monoclonal antibodies. Toxin prepared from different toxigenic strains could not be differentiated immunologically by tandem crossed immunelectrophoresis. Toxin, which was affinity purified from four of the strains and subsequently inactivated by formaldehyde, was cross‐protective when used for vaccination of mice before challenge with PMT. It is concluded that the toxin from toxigenic strains of P. multocida ssp. multocida must be very similar, if not identical.

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Section: Discussionmentioning

confidence: 79%

Characterization of Toxin from Different Strains of Pasteurella multocida Serotype A and D

Frandsen

Foged

Petersen

et al. 1991

Journal of Veterinary Medicine, Series B

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“…The toxin is also mitogenic for the following established cell lines, Rat-1 [3], BALB/c 3T3, NIH 3T3, 3T6 and human fibroblasts [1]. The toxin has been cloned, sequenced and expressed in Escherichia co# [4][5][6][7][8][9]. PMT is encoded as a 146 kDa singlechain bacterial protein, which shares partial homology with the multinucleating toxins, cytotoxic necrotising factors types 1 and 2 (CNF-1, CNF-2), produced by some strains of E. coli [10,11].…”

Section: Introductionmentioning

confidence: 99%

The potent mitogen Pasteurella multocida toxin is highly resistant to proteolysis but becomes susceptible at lysosomal pH

Smyth¹,

Pickersgill²,

Lax³

1995

FEBS Letters

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The susceptibility of the potent mitogen Pasteurella multocida toxin (PMT) to various proteases was investigated. PMT at a toxin to protease molar ratio of 1 : 1 was resistant to 8 of the 11 proteases tested after one hour. With longer incubation, PMT remained resistant to 7 proteases, and this correlated with a retention of biological activity, indicating that PMT might not require proteolytic cleavage at least until it bound to a cell receptor. Previous evidence had suggested that PMT is processed in the cell via an endosome or lysosome. We have shown that PMT became susceptible to proteolysis when the pH was lowered to 5 or below. This supports the previous suggestion that PMT is processed via a low pH compartment in the cell.

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“…The specificity of ToxA for toxigenic P. multocida further depends upon the assumption that nontoxigenic strains lack the toxA gene. Most available evidence is consistent with this assumption (4,5,9,10,11,14,15). However, a few reports present conflicting data suggesting that silent or incomplete copies of toxA may occasionally be found in nontoxigenic isolates (8,9,11).…”

Section: Discussionmentioning

confidence: 81%

“…Use of a ToxA-specific probe for identification of toxigenic stains of P. multocida further requires that homologous sequence be absent from phenotypically nontoxigenic strains. Available data suggest that this may generally be the case for nontoxigenic P. multocida (4,5,9,10,11,14,15). However, these studies included only a few isolates or isolates from a limited geographic region.…”

Section: Resultsmentioning

confidence: 99%

Two-Color Hybridization Assay for Simultaneous Detection of Bordetella bronchiseptica and Toxigenic Pasteurella multocida from Swine

Lee

Thomson

1998

J Clin Microbiol

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Bordetella bronchiseptica and toxigenic Pasteurella multocida are the etiologic agents of swine atrophic rhinitis. Methods currently used for their identification are time-consuming and suffer from a lack of sensitivity. We describe a colony lift-hybridization assay for detection of B. bronchiseptica and toxigenic P. multocida that can be performed with a single colony lift derived from a primary isolation plate without the need for pure subcultures of suspect bacteria. Membranes are hybridized simultaneously to probes derived from the B. bronchiseptica alcA gene and the P. multocida toxA gene. A multicolor development procedure permits sequential detection of bound probes. The assay was tested with 84 primary isolation plates generated from nasal swabs from swine with clinical signs of atrophic rhinitis. Comparison of the results from the colony lift-hybridization assay with those from conventional testing, based on a combination of colony morphology, biochemical reactions, mouse lethality, and enzyme-linked immunosorbent assay, indicated that the colony lift assay has superior sensitivity and comparable specificity. This technique has wide application for diagnostic and experimental studies.

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Cloning and expression of the dermonecrotic toxin gene of Pasteurella multocida ssp. multocida in Echerichia coli

Cited by 4 publications

References 4 publications

Characterization of Toxin from Different Strains of Pasteurella multocida Serotype A and D

Characterization of Toxin from Different Strains of Pasteurella multocida Serotype A and D

The potent mitogen Pasteurella multocida toxin is highly resistant to proteolysis but becomes susceptible at lysosomal pH

Two-Color Hybridization Assay for Simultaneous Detection of Bordetella bronchiseptica and Toxigenic Pasteurella multocida from Swine

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