title: Mutation analysis of pertussis toxin promoter.
abstract: The improving of the expression efficiency of a pertussis toxin (PT) promoter was believed to be a critical issue for the production of PT in acellular vaccine development. In this study, we have isolated a PT promoter region from the genome of a pertussis strain ATCC 9340. Based on the promoter sequence, a series of mutant PT promoters have been generated and subjected to in vitro gel shift analysis and in vivo reporter beta-galactosidase activity study. As compared with the wild type promoter, the mutation of the ribosome binding sequence or -10 element, to the respective consensus sequence derived from strong bacterial promoters, resulted in an enhancement of its interaction with two cellular proteins, and a slightly higher beta-galactosidase activity (1.3 fold). Whereas, the change of either upstream inverse repeats or 20-bp direct repeats to a certain complete repeat significantly promoted the formation of another DNA-protein-complex, and exhibited an 1.8 fold beta-galactosidase activity. These findings would have provided a mutation target for making a more efficient PT-production pertussis strain.
authors.affiliation: Division of Biologics Quality Control, National Institute of Preventive Medicine, Taipei, Taiwan, ROC.
authors.forename: Y Y
authors.initials: YY
authors.lastname: Wo
authors.forename: S C
authors.initials: SC
authors.lastname: Chiu
authors.forename: C H
authors.initials: CH
authors.lastname: Lu
mesh_terms: D001483:Base Sequence; D008969:Molecular Sequence Data; D009154:Mutation; D037342:Pertussis Toxin; D011401:Promoter Regions, Genetic; D010566:Virulence Factors, Bordetella / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin