title: Isolation of an asporogenic (spoOA) protective antigen-producing strain of Bacillus anthracis.
abstract: We found that Congo red agar allows identification of sporulation-deficient Bacillus anthracis. Using Congo red agar, we isolated an asporogenic derivative of the protective antigen-producing strain B. anthracis delta Sterne-1(pPA102). Polymerase chain reaction and Southern hybridization analyses of DNA from the asporogenic mutant revealed that a deletion was present in spoOA, an essential gene for the initiation of sporulation. The deletion also encompassed the spoIVB homologue and a portion of the recN homologue. The avirulent spoOA strain delta Sterne-1(pPA102)CR4 is suitable for the safe production of protective antigen without endospore contamination of the vaccine production facility.
authors.affiliation: Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Frederick, MD 21702-5011, USA. worsham@ncifcrf.gov
authors.forename: P L
authors.initials: PL
authors.lastname: Worsham
authors.forename: M R
authors.initials: MR
authors.lastname: Sowers
mesh_terms: D000362:Agar; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000276:immunology*; D001408:Bacillus anthracis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification* / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics*; D001428:Bacterial Vaccines / Q000276:immunology; D015139:Blotting, Southern; D002874:Chromosome Mapping; D016386:Chromosome Walking; D003224:Congo Red; D017353:Gene Deletion; D005798:Genes, Bacterial; D016133:Polymerase Chain Reaction / Q000379:methods; D013171:Spores, Bacterial / Q000502:physiology; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; C080950:Spo0A protein, Bacillus subtilis; D014157:Transcription Factors; D003224:Congo Red; D000362:Agar