title: Purification and characterization of neurotoxin complex from a dual toxin gene containing Clostridium Botulinum Strain PS-5.
abstract: Botulinum neurotoxins are produced as a toxin complex (TC) which consists of neurotoxin (NT) and neurotoxin associated proteins. The characterization of NT in its native state is an essential step for developing diagnostics and therapeutic countermeasures against botulism. The presence of NT genes was validated by PCR amplification of toxin specific fragments from genomic DNA of Clostridium botulinum strain PS-5 which indicated the presence of both serotype A and B genes on PS-5 genome. Further, TC was purified and characterized by Western blotting, Digoxin-enzyme linked immunosorbent assay, endopeptidase activity assay, and Liquid chromatography-Mass spectrometry. The data showed the presence of serotype A specific neurotoxin. Based on the analysis of neurotoxin genes and characterization of TC, PS-5 strain appears as a serotype A (B) strain of C. botulinum which produces only serotype A specific TC in the cell culture medium.
authors.affiliation: Divisions of Microbiology, Center for Food Safety and Applied Nutrition, Food and Drug Administration, College Park, MD 20740, USA. ajay.singh@fda.hhs.gov
authors.forename: Ajay K
authors.initials: AK
authors.lastname: Singh
authors.forename: Amita
authors.initials: A
authors.lastname: Sachdeva
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Degrasse
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Croley
authors.forename: Larry H
authors.initials: LH
authors.lastname: Stanker
authors.forename: David
authors.initials: D
authors.lastname: Hodge
authors.forename: Shashi K
authors.initials: SK
authors.lastname: Sharma
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D015153:Blotting, Western; D002645:Chickens; D003014:Clostridium botulinum / Q000737:chemistry* / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005798:Genes, Bacterial / Q000235:genetics; D006386:Hemagglutination Tests; D046912:Multiprotein Complexes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D009498:Neurotoxins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D016133:Polymerase Chain Reaction
chemical_list: D001426:Bacterial Proteins; D046912:Multiprotein Complexes; D009498:Neurotoxins