{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T15:56:03Z","timestamp":1672934163655},"reference-count":19,"publisher":"Microbiology Society","issue":"5","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2009,5,1]]},"abstract":"<jats:p>Haemolysin BL (HBL) of <jats:italic>Bacillus cereus<\/jats:italic>, listeriolysin O (LLO) of <jats:italic>Listeria monocytogenes<\/jats:italic> and enterotoxin B (SEB) of <jats:italic>Staphylococcus aureus<\/jats:italic> are among the major toxin components contributing to the pathogenicity of these organisms in foodborne illnesses. In this study, an intergenus non-toxic multidomain fusion protein (r-HLE) was generated with specificity for HBL, LLO and SEB. The fusion gene (<jats:italic>r-hle<\/jats:italic>) comprising the conserved regions of <jats:italic>hblD<\/jats:italic> and the <jats:italic>hly<\/jats:italic> and <jats:italic>entB<\/jats:italic> genes was codon-optimized for expression in <jats:italic>Escherichia coli<\/jats:italic> and encoded a 50\u2005kDa recombinant multidomain chimeric protein (r-HLE). Hyperimmune antiserum raised against r-HLE specifically reacted with the L<jats:sub>1<\/jats:sub> (38\u2005kDa) component of the HBL complex of <jats:italic>B. cereus<\/jats:italic>, LLO (58\u2005kDa) of <jats:italic>L. monocytogenes<\/jats:italic> and SEB (28\u2005kDa) of <jats:italic>S. aureus<\/jats:italic> during Western blot analysis when tested on standard strains. During testing on isolates, the antiserum again identified the appropriate toxin molecules and was highly specific to the relevant bacterial species. The antigenicity of the SEB component of the r-HLE protein was also confirmed using a commercially available TECRA kit. The described procedure of creating a single antigenic molecule carrying components of three different toxins whilst still retaining the original antigenic determinants of individual toxins will be highly advantageous in the development of rapid, reliable and cost-effective immunoassays.<\/jats:p>","DOI":"10.1099\/jmm.0.007658-0","type":"journal-article","created":{"date-parts":[[2009,4,15]],"date-time":"2009-04-15T17:38:13Z","timestamp":1239817093000},"page":"577-583","source":"Crossref","is-referenced-by-count":4,"title":["Construction of a recombinant intergenus multidomain chimeric protein for simultaneous expression of haemolysin BL of Bacillus cereus, listeriolysin O of Listeria monocytogenes and enterotoxin B of Staphylococcus aureus"],"prefix":"10.1099","volume":"58","author":[{"given":"T. D. 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