Project description:Comparative genomic hybridizations were performed to compare bont/A1 strains with an A2-like toxin gene cluster organization to the genome sequenced strain, C. botulinum ATCC 3502. Keywords: comparative genomic hybridization
Project description:Strains were differentiated on the basis of hybridization to probes representing strain variable regions in C. botulinum strain ATCC 3502. Probes for selected genes (eg. toxin genes) were also featured on the microarray to allow detection of other serotypes/subtypes.
Project description:Strains were differentiated on the basis of hybridization to probes representing strain variable regions in C. botulinum strain ATCC 3502. Probes for selected genes (eg. toxin genes) were also featured on the microarray to allow detection of other serotypes/subtypes.
Project description:Comparative genomic hybridizations were performed to compare bont/A1 strains with an A2-like toxin gene cluster organization to the genome sequenced strain, C. botulinum ATCC 3502. Keywords: comparative genomic hybridization For each hybridization, the reference strain (C. botulinum ATCC 3502) was labeled with Cy5 and each test strain was labeled with Cy3.
Project description:Clostridium botulinum is a heterogeneous Gram-positive species that comprises four genetically and physiologically distinct groups of bacteria that share the ability to produce botulinum neurotoxin, the most poisonous toxin known to man, and the causative agent of botulism, a severe disease of humans and animals. We report here the complete genome sequence of a representative of Group I (proteolytic) C. botulinum (strain Hall A, ATCC 3502). The genome consists of a chromosome (3,886,916 bp) and a plasmid (16,344 bp) which carry 3,650 and 19 predicted genes, respectively. Consistent with the proteolytic phenotype of this strain, the genome harbours a large number of genes encoding secreted proteases and enzymes involved in uptake and metabolism of amino acids. The genome also reveals a hitherto unknown ability of C. botulinum to degrade chitin. There is a significant lack of recently acquired DNA, indicating a stable genomic content, in strong contrast to the fluid genome of C. difficile, which can form longer-term relationships with its host. Overall, the genome indicates that C. botulinum is adapted to a saprophytic lifestyle both in soil and aquatic environments. This pathogen relies on its toxin to rapidly kill a wide range of prey species, and to gain access to nutrient sources, it releases a large number of extracellular enzymes to soften and destroy rotting or decayed tissues.
Project description:In this study, we identified the O-glycosylation sites in deletion constructs of recombinant Clostridium botulinum flagellin (CbFla) expressed from the E. coli strain, EV136 (K1:K12 strains engineered by Prof. Eric Vimr’s group to accumulate CMP-sialic acid in the cytosol). The recombinant flagellins were expressed with or without co-expression of motility associated factor (Maf) (flagellin nonulosonic acid glycosyltransferase) in these strains, purified and subjected to MS/MS analysis at Taplin Biological Mass Spectrometry facility. We also studied the O-glycosylation of recombinant Clostridium botulinum flagellin (CbFla) co-expressed with Geobacillus kaustophilus Maf. We also studied the O-glycosylation of flagellin chimeras made by grafting mini-flagellin sequences on to an unrelated protein with an F-type lectin domain from Streptosporangium roseum SrNaFLD.
Project description:Comparative genomic hybridization microarrays featuring overlapping probes spanning the entire C. botulinum type A1 strain ATCC 3502 genome were used to identify regions whose presence are variable among a diverse panel of type A C. botulinum strains.
Project description:The purpose of this study was to compare the gene content of several group II C. botulinum strains to determine the utility of DNA microarrays for subtyping this organism. Group II type B, E, and F C. botulinum strains were compared in a study to examine the gene content of a unique type E strain (CDC66177).