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. 1983 Apr;154(1):419–428. doi: 10.1128/jb.154.1.419-428.1983

Diversity of locations for Bacillus thuringiensis crystal protein genes.

J W Kronstad, H E Schnepf, H R Whiteley
PMCID: PMC217475  PMID: 6833183

Abstract

The location of crystal protein genes in 22 crystalliferous Bacillus thuringiensis strains representing 14 subspecies was investigated by hybridization of an intragenic restriction fragment from a cloned crystal protein gene to whole plasmid preparations. Hybridization was found to a single plasmid in eight strains, to more than one plasmid in seven strains, and to one or both of two large, unresolved plasmids in two strains. The sizes of the hybridized plasmids ranged from 33 to over 150 megadaltons. In one additional subspecies, hybridization was only to linear DNA fragments, suggesting a chromosomal crystal protein gene, and for four other subspecies, not reported to be toxic to lepidopteran insects, no hybridization was found to either plasmids or to total cell DNA. Hybridization to restriction digests of plasmids and total cell DNA of several strains of subspecies thuringiensis and kurstaki revealed that all homology to the cloned crystal protein gene was plasmid associated and that several of these strains contained multiple regions of homology, implying the presence of multiple crystal protein genes.

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Selected References

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  1. Aronson A. I., Angelo N., Holt S. C. Regulation of extracellular protease production in Bacillus cereus T: characterization of mutants producing altered amounts of protease. J Bacteriol. 1971 Jun;106(3):1016–1025. doi: 10.1128/jb.106.3.1016-1025.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aronson A. I., Tyrell D. J., Fitz-James P. C., Bulla L. A., Jr Relationship of the syntheses of spore coat protein and parasporal crystal protein in Bacillus thuringiensis. J Bacteriol. 1982 Jul;151(1):399–410. doi: 10.1128/jb.151.1.399-410.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Calabrese D. M., Nickerson K. W., Lane L. C. A comparison of protein crystal subunit sizes in Bacillus thuringiensis. Can J Microbiol. 1980 Aug;26(8):1006–1010. doi: 10.1139/m80-170. [DOI] [PubMed] [Google Scholar]
  5. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  6. Dulmage H. T. Production of -endotoxin by eighteen isolates of Bacillus thuringiensis, serotype 3, in 3 fermentation media. J Invertebr Pathol. 1971 Nov;18(3):353–358. doi: 10.1016/0022-2011(71)90037-1. [DOI] [PubMed] [Google Scholar]
  7. George J., Chirikjian J. G. Sequence-specific endonuclease BamHI: relaxation of sequence recognition. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2432–2436. doi: 10.1073/pnas.79.8.2432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. González J. M., Jr, Brown B. J., Carlton B. C. Transfer of Bacillus thuringiensis plasmids coding for delta-endotoxin among strains of B. thuringiensis and B. cereus. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6951–6955. doi: 10.1073/pnas.79.22.6951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. González J. M., Jr, Carlton B. C. Patterns of plasmid DNA in crystalliferous and acrystalliferous strains of Bacillus thuringiensis. Plasmid. 1980 Jan;3(1):92–98. doi: 10.1016/s0147-619x(80)90038-4. [DOI] [PubMed] [Google Scholar]
  10. González J. M., Jr, Dulmage H. T., Carlton B. C. Correlation between specific plasmids and delta-endotoxin production in Bacillus thuringiensis. Plasmid. 1981 May;5(3):352–365. doi: 10.1016/0147-619x(81)90010-x. [DOI] [PubMed] [Google Scholar]
  11. Held G. A., Bulla L. A., Jr, Ferrari E., Hoch J., Aronson A. I., Minnich S. A. Cloning and localization of the lepidopteran protoxin gene of Bacillus thuringiensis subsp. kurstaki. Proc Natl Acad Sci U S A. 1982 Oct;79(19):6065–6069. doi: 10.1073/pnas.79.19.6065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Howley P. M., Israel M. A., Law M. F., Martin M. A. A rapid method for detecting and mapping homology between heterologous DNAs. Evaluation of polyomavirus genomes. J Biol Chem. 1979 Jun 10;254(11):4876–4883. [PubMed] [Google Scholar]
  13. Klier A., Fargette F., Ribier J., Rapoport G. Cloning and expression of the crystal protein genes from Bacillus thuringiensis strain berliner 1715. EMBO J. 1982;1(7):791–799. doi: 10.1002/j.1460-2075.1982.tb01249.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Krywienczyk J., Dulmage H. T., Fast P. G. Occurrence of two serologically distinct groups within Bacillus thuringiensis serotype 3 ab var. kurstaki. J Invertebr Pathol. 1978 May;31(3):372–375. doi: 10.1016/0022-2011(78)90232-x. [DOI] [PubMed] [Google Scholar]
  15. Maniatis T., Hardison R. C., Lacy E., Lauer J., O'Connell C., Quon D., Sim G. K., Efstratiadis A. The isolation of structural genes from libraries of eucaryotic DNA. Cell. 1978 Oct;15(2):687–701. doi: 10.1016/0092-8674(78)90036-3. [DOI] [PubMed] [Google Scholar]
  16. Messing J., Crea R., Seeburg P. H. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. doi: 10.1093/nar/9.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schnepf H. E., Whiteley H. R. Cloning and expression of the Bacillus thuringiensis crystal protein gene in Escherichia coli. Proc Natl Acad Sci U S A. 1981 May;78(5):2893–2897. doi: 10.1073/pnas.78.5.2893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith H. O. Recovery of DNA from gels. Methods Enzymol. 1980;65(1):371–380. doi: 10.1016/s0076-6879(80)65048-4. [DOI] [PubMed] [Google Scholar]
  19. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  20. Spizizen J. TRANSFORMATION OF BIOCHEMICALLY DEFICIENT STRAINS OF BACILLUS SUBTILIS BY DEOXYRIBONUCLEATE. Proc Natl Acad Sci U S A. 1958 Oct 15;44(10):1072–1078. doi: 10.1073/pnas.44.10.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stahly D. P., Dingman D. W., Bulla L. A., Jr, Aronson A. I. Possible origin and function of the parasporal crystal in Bacillus thuringiensis. Biochem Biophys Res Commun. 1978 Oct 16;84(3):581–588. doi: 10.1016/0006-291x(78)90745-3. [DOI] [PubMed] [Google Scholar]
  22. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. White F. F., Nester E. W. Hairy root: plasmid encodes virulence traits in Agrobacterium rhizogenes. J Bacteriol. 1980 Mar;141(3):1134–1141. doi: 10.1128/jb.141.3.1134-1141.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wong H. C., Schnepf H. E., Whiteley H. R. Transcriptional and translational start sites for the Bacillus thuringiensis crystal protein gene. J Biol Chem. 1983 Feb 10;258(3):1960–1967. [PubMed] [Google Scholar]
  25. Yamamoto T., McLaughlin R. E. Isolation of a protein from the parasporal crystal of Bacillus thuringiensis var. Kurstaki toxic to the mosquito larva, Aedes taeniorhynchus. Biochem Biophys Res Commun. 1981 Nov 30;103(2):414–421. doi: 10.1016/0006-291x(81)90468-x. [DOI] [PubMed] [Google Scholar]

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