ORBi: Lebrun Maud - Purification of the recombinant beta2 toxin (CPB2) from an enterotoxaemic bovine Clostridium perfringens strain and production of a specific immune serum

Reference : Purification of the recombinant beta2 toxin (CPB2) from an enterotoxaemic bovine Clostri...


	Document type :	Scientific journals : Article
	Discipline(s) :	Life sciences : Microbiology Life sciences : Biochemistry, biophysics & molecular biology Life sciences : Biotechnology Life sciences : Veterinary medicine & animal health
	To cite this reference:	http://hdl.handle.net/2268/26817

Title :	Purification of the recombinant beta2 toxin (CPB2) from an enterotoxaemic bovine Clostridium perfringens strain and production of a specific immune serum
Language :	English
Author, co-author :	Lebrun, Maud [Université de Liège - ULG > Département des Maladies Infectieuses et Parasitaires > Bactériologie et pathologie des maladies bactériennes > >]
	Filée, Patrice [Université de Liège - ULg > > Centre d'ingénierie des protéines >]
	Galleni, Moreno [Université de Liège - ULg > Département des sciences de la vie > Macromolécules biologiques >]
	Mainil, Jacques [Université de Liège - ULg > Département des maladies infectieuses et parasitaires > Bactériologie et pathologie des maladies bactériennes >]
	Linden, Annick [Université de Liège - ULg > Département des maladies infectieuses et parasitaires > Santé et pathologies de la faune sauvage >]
	Taminiau, Bernard [Université de Liège - ULg > Département de sciences des denrées alimentaires > Microbiologie des denrées alimentaires >]
Publication date :	Sep-2007
Journal title :	Protein Expression & Purification
Publisher :	Academic Press
Volume :	55
Issue/season :	1
Pages :	119-131
Audience :	International
ISSN :	1046-5928
e-ISSN :	1096-0279
City :	Orlando
Country :	FL
Keywords :	[en] Clostridium perfringens ; beta2-toxin ; cloning ; expression ; Bacillus subtilis ; Escherichia coli
Abstract :	[en] Overgrowth of Clostridium perfringens clones with production of one or more of its toxin(s) results in diverse digestive and systemic pathologies in human and animals, such as cattle enterotoxaemia. The so-called beta2 toxin (CPB2) is the most recently described major toxin produced by C perfringens. In this study, the cpb2 ORF (cpb2FM) from a cattle C perfringens-associated enterotoxaemia was cloned and sequenced. The cpb2FM and its deduced nucleotide sequence clearly corresponded to the epb2 allele considered as "consensus" and not to "atypical" allele, despite its "non-porcine" origin. Expression assays of the recombinant toxin CPB2FM were performed in Escherichia coli and Bacillus subtilis with the expression vector pBLTS72, and by genomic integration by double recombination in B. subtilis. Highest level of production was obtained with the expression vector in B. subtilis 168 strain. The recombinant CPB2FM protein was purified and a specific rabbit polyclonal antiserum was produced. Polyclonal antibodies could detect CPB2 production in supernatants of C. perfringens from enterotoxaemic cattle. (C) 2007 Elsevier Inc. All rights reserved.
Permalink :	http://hdl.handle.net/2268/26817
	also: http://hdl.handle.net/2268/104907
DOI :	10.1016/j.pep.2007.04.021

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