Evidence for plasmid-associated crystal toxin production in Bacillus thuringiensis subsp. israelensis |
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Authors: | Robert M Faust Kazunori Abe Gary A Held Toshihiko Iizuka Lee A Bulla Carol L Meyers |
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Institution: | 1. Insect Pathology Laboratory, ARS-S&E-USDA, Beltsville, Maryland 20705 USA;2. Department of Food and Nutrition, Seinan Jogakuin Junior College, Kitakyushu 803, Japan;3. Department of Bacteriology and Biochemistry, University of Idaho, Moscow, Idaho 83843 U.S.S.R.;4. Laboratory of Sericology, Hokkaido University, Sapporo 060, Japan;5. Department of Entomology, University of Maryland, College Park, Maryland 20740 USA |
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Abstract: | Three crystalliferous (Cry+) strains of Bacillus thuringiensis subsp. israelensis (serotype 14) that produce parasporal protein crystals toxic to dipteran larvae and several acrystalliferous (Cry?) mutants, either induced or spontaneously derived from a single Cry+ parent, were examined for the presence of covalently closed circular (CCC) DNA in attempts to correlate toxin production with the presence of a specific plasmid. The plasmid profiles of both Cry+ and Cry? variants were analyzed by both a cleared lysate- and a modified Eckhardt lysateelectrophoresis technique. All of the Cry? mutants derived from the Cry+ parental strain had lost a 4.0- to 4.4-megadalton (Mdal) plasmid. Bioassay data confirmed loss of toxin production by the Cry? variants. All three Cry+ strains, including the parent of the Cry? strains, contained CCC plasmids DNAs of the following approximate molecular weights: 4.0 to 4.4, 5.2 to 6.0, and 11.4 to 13.0 Mdal. One Cry+ strain contained an additional CCC plasmid of 6.7 to 7.2 Mdal. The plasmid patterns for several Cry? derivatives differed in other respects from the pattern for their parent strain. The various Cry+ and Cry? strains could be distinguished either by phenotypical differences in antibiotic sensitivity, crystal production, and toxicity, or by differences in their plasmid profiles. |
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Keywords: | To whom all correspondence should be addressed: USDA S&E Insect Pathology Laboratory Rm 214 Bldg 011A Biosci Agr Res Center—West Beltsville Md 20705 |
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