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. 1987 Dec;53(12):2808–2814. doi: 10.1128/aem.53.12.2808-2814.1987

Use of oligonucleotide probes to study the relatedness of delta-endotoxin genes among Bacillus thuringiensis subspecies and strains.

G Prefontaine 1, P Fast 1, P C Lau 1, M A Hefford 1, Z Hanna 1, R Brousseau 1
PMCID: PMC204203  PMID: 2829720

Abstract

Fifteen Bacillus thuringiensis strains representing 13 serotypes were screened with five oligodeoxyribonucleotide probes specific for certain regions of two published sequences and one unpublished sequence of B. thuringiensis delta-endotoxin genes. Of the 15 cultures, 14 hybridized with at least one probe; the B. thuringiensis subsp. thompsoni strain alone did not hybridize. Two B. thuringiensis subsp. kurstaki strains of commercial interest, HD-1 and NRD-12, were found to be so closely related as to be indistinguishable with this technique; the same situation was found with strains from B. thuringiensis subspp. dendrolimus and sotto. Five strains were identified as probably containing only one endotoxin gene. A probe specific for the gene from the B. thuringiensis subsp. kurstaki HD-73 strain hybridized to only 3 of the 15 cultures tested. The hybridization data suggest that the DNA sequences coding for the C-terminal region of the endotoxin protein are as well conserved as those coding for the N-terminal toxic portion.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adang M. J., Staver M. J., Rocheleau T. A., Leighton J., Barker R. F., Thompson D. V. Characterized full-length and truncated plasmid clones of the crystal protein of Bacillus thuringiensis subsp. kurstaki HD-73 and their toxicity to Manduca sexta. Gene. 1985;36(3):289–300. doi: 10.1016/0378-1119(85)90184-2. [DOI] [PubMed] [Google Scholar]
  2. Aronson A. I., Beckman W., Dunn P. Bacillus thuringiensis and related insect pathogens. Microbiol Rev. 1986 Mar;50(1):1–24. doi: 10.1128/mr.50.1.1-24.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bouché J. P. The effect of spermidine on endonuclease inhibition by agarose contaminants. Anal Biochem. 1981 Jul 15;115(1):42–45. doi: 10.1016/0003-2697(81)90519-4. [DOI] [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. Chapman J. S., Carlton B. C. Conjugal plasmid transfer in Bacillus thuringiensis. Basic Life Sci. 1985;30:453–467. doi: 10.1007/978-1-4613-2447-8_33. [DOI] [PubMed] [Google Scholar]
  6. Chou P. Y., Fasman G. D. Prediction of protein conformation. Biochemistry. 1974 Jan 15;13(2):222–245. doi: 10.1021/bi00699a002. [DOI] [PubMed] [Google Scholar]
  7. Dalbadie-McFarland G., Cohen L. W., Riggs A. D., Morin C., Itakura K., Richards J. H. Oligonucleotide-directed mutagenesis as a general and powerful method for studies of protein function. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6409–6413. doi: 10.1073/pnas.79.21.6409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Debro L., Fitz-James P. C., Aronson A. Two different parasporal inclusions are produced by Bacillus thuringiensis subsp. finitimus. J Bacteriol. 1986 Jan;165(1):258–268. doi: 10.1128/jb.165.1.258-268.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Geiser M., Schweitzer S., Grimm C. The hypervariable region in the genes coding for entomopathogenic crystal proteins of Bacillus thuringiensis: nucleotide sequence of the kurhd1 gene of subsp. kurstaki HD1. Gene. 1986;48(1):109–118. doi: 10.1016/0378-1119(86)90357-4. [DOI] [PubMed] [Google Scholar]
  10. Höfte H., de Greve H., Seurinck J., Jansens S., Mahillon J., Ampe C., Vandekerckhove J., Vanderbruggen H., van Montagu M., Zabeau M. Structural and functional analysis of a cloned delta endotoxin of Bacillus thuringiensis berliner 1715. Eur J Biochem. 1986 Dec 1;161(2):273–280. doi: 10.1111/j.1432-1033.1986.tb10443.x. [DOI] [PubMed] [Google Scholar]
  11. Kronstad J. W., Schnepf H. E., Whiteley H. R. Diversity of locations for Bacillus thuringiensis crystal protein genes. J Bacteriol. 1983 Apr;154(1):419–428. doi: 10.1128/jb.154.1.419-428.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kronstad J. W., Whiteley H. R. Inverted repeat sequences flank a Bacillus thuringiensis crystal protein gene. J Bacteriol. 1984 Oct;160(1):95–102. doi: 10.1128/jb.160.1.95-102.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kronstad J. W., Whiteley H. R. Three classes of homologous Bacillus thuringiensis crystal-protein genes. Gene. 1986;43(1-2):29–40. doi: 10.1016/0378-1119(86)90005-3. [DOI] [PubMed] [Google Scholar]
  14. Lereclus D., Ribier J., Klier A., Menou G., Lecadet M. M. A transposon-like structure related to the delta-endotoxin gene of Bacillus thuringiensis. EMBO J. 1984 Nov;3(11):2561–2567. doi: 10.1002/j.1460-2075.1984.tb02174.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Meinkoth J., Wahl G. Hybridization of nucleic acids immobilized on solid supports. Anal Biochem. 1984 May 1;138(2):267–284. doi: 10.1016/0003-2697(84)90808-x. [DOI] [PubMed] [Google Scholar]
  16. Oeda K., Oshie K., Shimizu M., Nakamura K., Yamamoto H., Nakayama I., Ohkawa H. Nucleotide sequence of the insecticidal protein gene of Bacillus thuringiensis strain aizawai IPL7 and its high-level expression in Escherichia coli. Gene. 1987;53(1):113–119. doi: 10.1016/0378-1119(87)90098-9. [DOI] [PubMed] [Google Scholar]
  17. Sanchez-Pescador R., Urdea M. S. Use of unpurified synthetic deoxynucleotide primers for rapid dideoxynucleotide chain termination sequencing. DNA. 1984 Aug;3(4):339–343. doi: 10.1089/dna.1.1984.3.339. [DOI] [PubMed] [Google Scholar]
  18. Schnepf H. E., Whiteley H. R. Delineation of a toxin-encoding segment of a Bacillus thuringiensis crystal protein gene. J Biol Chem. 1985 May 25;260(10):6273–6280. [PubMed] [Google Scholar]
  19. Shibano Y., Yamagata A., Nakamura N., Iizuka T., Sugisaki H., Takanami M. Nucleotide sequence coding for the insecticidal fragment of the Bacillus thuringiensis crystal protein. Gene. 1985;34(2-3):243–251. doi: 10.1016/0378-1119(85)90133-7. [DOI] [PubMed] [Google Scholar]
  20. Thorne L., Garduno F., Thompson T., Decker D., Zounes M., Wild M., Walfield A. M., Pollock T. J. Structural similarity between the lepidoptera- and diptera-specific insecticidal endotoxin genes of Bacillus thuringiensis subsp. "kurstaki" and "israelensis". J Bacteriol. 1986 Jun;166(3):801–811. doi: 10.1128/jb.166.3.801-811.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wabiko H., Raymond K. C., Bulla L. A., Jr Bacillus thuringiensis entomocidal protoxin gene sequence and gene product analysis. DNA. 1986 Aug;5(4):305–314. doi: 10.1089/dna.1986.5.305. [DOI] [PubMed] [Google Scholar]

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