Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1988 Aug;170(8):3575–3583. doi: 10.1128/jb.170.8.3575-3583.1988

A Bacillus thuringiensis subsp. israelensis gene encoding a 125-kilodalton larvicidal polypeptide is associated with inverted repeat sequences.

C Bourgouin 1, A Delécluse 1, J Ribier 1, A Klier 1, G Rapoport 1
PMCID: PMC211331  PMID: 2841292

Abstract

A gene encoding a 125-kilodalton (kDa) mosquitocidal delta-endotoxin was cloned from the 72-MDa resident plasmid of Bacillus thuringiensis subsp. israelensis. This gene is similar in its 3' region to the gene encoding the 135-kDa protein previously cloned (C. Bourgouin, A. Klier, and G. Rapoport, Mol. Gen. Genet. 205:390-397, 1986). Escherichia coli recombinant clones harboring the 125-kDa gene were toxic to larvae of the three mosquito species Aedes aegypti, Anopheles stephensi, and Culex pipiens. In addition, the B. thuringiensis subsp. israelensis DNA fragment carrying the 125-kDa protein gene contains two sets of inverted repeat sequences, identified either by the S1 nuclease method or by electron microscopic observation. The structural organization of inverted repeat sequences and of the 125-kDa gene was analyzed and suggests that this B. thuringiensis subsp. israelensis delta-endotoxin gene is located within a transposable element.

Full text

PDF
3575

Images in this article

3577Image
on p.3577
3578Image
on p.3578
3578Image
on p.3578
3579Image
on p.3579
3580Image
on p.3580

Selected References

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

  1. Alwine J. C., Kemp D. J., Parker B. A., Reiser J., Renart J., Stark G. R., Wahl G. M. Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper. Methods Enzymol. 1979;68:220–242. doi: 10.1016/0076-6879(79)68017-5. [DOI] [PubMed] [Google Scholar]
  2. Angsuthanasombat C., Chungjatupornchai W., Kertbundit S., Luxananil P., Settasatian C., Wilairat P., Panyim S. Cloning and expression of 130-kd mosquito-larvicidal delta-endotoxin gene of Bacillus thuringiensis var. Israelensis in Escherichia coli. Mol Gen Genet. 1987 Jul;208(3):384–389. doi: 10.1007/BF00328128. [DOI] [PubMed] [Google Scholar]
  3. Betley M. J., Miller V. L., Mekalanos J. J. Genetics of bacterial enterotoxins. Annu Rev Microbiol. 1986;40:577–605. doi: 10.1146/annurev.mi.40.100186.003045. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Bourgouin C., Klier A., Rapoport G. Characterization of the genes encoding the haemolytic toxin and the mosquitocidal delta-endotoxin of Bacillus thuringiensis israelensis. Mol Gen Genet. 1986 Dec;205(3):390–397. doi: 10.1007/BF00338072. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Earp D. J., Ellar D. J. Bacillus thuringiensis var. morrisoni strain PG14: nucleotide sequence of a gene encoding a 27kDa crystal protein. Nucleic Acids Res. 1987 Apr 24;15(8):3619–3619. doi: 10.1093/nar/15.8.3619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Garduno F., Thorne L., Walfield A. M., Pollock T. J. Structural relatedness between mosquitocidal endotoxins of Bacillus thuringiensis subsp. israelensis. Appl Environ Microbiol. 1988 Jan;54(1):277–279. doi: 10.1128/aem.54.1.277-279.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. González J. M., Jr, Carlton B. C. A large transmissible plasmid is required for crystal toxin production in Bacillus thuringiensis variety israelensis. Plasmid. 1984 Jan;11(1):28–38. doi: 10.1016/0147-619x(84)90004-0. [DOI] [PubMed] [Google Scholar]
  10. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  11. Hurley J. M., Bulla L. A., Jr, Andrews R. E., Jr Purification of the mosquitocidal and cytolytic proteins of Bacillus thuringiensis subsp. israelensis. Appl Environ Microbiol. 1987 Jun;53(6):1316–1321. doi: 10.1128/aem.53.6.1316-1321.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hurley J. M., Lee S. G., Andrews R. E., Jr, Klowden M. J., Bulla L. A., Jr Separation of the cytolytic and mosquitocidal proteins of Bacillus thuringiensis subsp. israelensis. Biochem Biophys Res Commun. 1985 Jan 31;126(2):961–965. doi: 10.1016/0006-291x(85)90279-7. [DOI] [PubMed] [Google Scholar]
  13. Kleckner N. Transposable elements in prokaryotes. Annu Rev Genet. 1981;15:341–404. doi: 10.1146/annurev.ge.15.120181.002013. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lee S. G., Eckblad W., Bulla L. A., Jr Diversity of protein inclusion bodies and identification of mosquitocidal protein in Bacillus thuringiensis subsp. israelensis. Biochem Biophys Res Commun. 1985 Jan 31;126(2):953–960. doi: 10.1016/0006-291x(85)90278-5. [DOI] [PubMed] [Google Scholar]
  18. Lereclus D., Lecadet M. M., Ribier J., Dedonder R. Molecular relationships among plasmids of Bacillus thuringiensis: conserved sequences through 11 crystalliferous strains. Mol Gen Genet. 1982;186(3):391–398. doi: 10.1007/BF00729459. [DOI] [PubMed] [Google Scholar]
  19. Lereclus D., Mahillon J., Menou G., Lecadet M. M. Identification of Tn4430, a transposon of Bacillus thuringiensis functional in Escherichia coli. Mol Gen Genet. 1986 Jul;204(1):52–57. doi: 10.1007/BF00330186. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Mahillon J., Seurinck J., Delcour J., Zabeau M. Cloning and nucleotide sequence of different iso-IS231 elements and their structural association with the Tn4430 transposon in Bacillus thuringiensis. Gene. 1987;51(2-3):187–196. doi: 10.1016/0378-1119(87)90307-6. [DOI] [PubMed] [Google Scholar]
  22. Mahillon J., Seurinck J., van Rompuy L., Delcour J., Zabeau M. Nucleotide sequence and structural organization of an insertion sequence element (IS231) from Bacillus thuringiensis strain berliner 1715. EMBO J. 1985 Dec 30;4(13B):3895–3899. doi: 10.1002/j.1460-2075.1985.tb04163.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McLean K. M., Whiteley H. R. Expression in Escherichia coli of a cloned crystal protein gene of Bacillus thuringiensis subsp. israelensis. J Bacteriol. 1987 Mar;169(3):1017–1023. doi: 10.1128/jb.169.3.1017-1023.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ohtsubo H., Ohtsubo E. Isolation of inverted repeat sequences, including IS1, IS2, and IS3, in Escherichia coli plasmids. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2316–2320. doi: 10.1073/pnas.73.7.2316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pfannenstiel M. A., Couche G. A., Ross E. J., Nickerson K. W. Immunological relationships among proteins making up the Bacillus thuringiensis subsp. israelensis crystalline toxin. Appl Environ Microbiol. 1986 Oct;52(4):644–649. doi: 10.1128/aem.52.4.644-649.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  27. Sanchis V., Lereclus D., Menou G., Chaufaux J., Lecadet M. M. Multiplicity of delta-endotoxin genes with different insecticidal specificities in Bacillus thuringiensis aizawai 7.29. Mol Microbiol. 1988 May;2(3):393–404. doi: 10.1111/j.1365-2958.1988.tb00044.x. [DOI] [PubMed] [Google Scholar]
  28. Sekar V., Carlton B. C. Molecular cloning of the delta-endotoxin gene of Bacillus thuringiensis var. israelensis. Gene. 1985;33(2):151–158. doi: 10.1016/0378-1119(85)90089-7. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Thomas W. E., Ellar D. J. Bacillus thuringiensis var israelensis crystal delta-endotoxin: effects on insect and mammalian cells in vitro and in vivo. J Cell Sci. 1983 Mar;60:181–197. doi: 10.1242/jcs.60.1.181. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Tungpradubkul S., Settasatien C., Panyim S. The complete nucleotide sequence of a 130 kDa mosquito-larvicidal delta-endotoxin gene of Bacillus thuringiensis var. israelensis. Nucleic Acids Res. 1988 Feb 25;16(4):1637–1638. doi: 10.1093/nar/16.4.1637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Waalwijk C., Dullemans A. M., van Workum M. E., Visser B. Molecular cloning and the nucleotide sequence of the Mr 28 000 crystal protein gene of Bacillus thuringiensis subsp. israelensis. Nucleic Acids Res. 1985 Nov 25;13(22):8207–8217. doi: 10.1093/nar/13.22.8207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Ward E. S., Ellar D. J. Bacillus thuringiensis var. israelensis delta-endotoxin. Nucleotide sequence and characterization of the transcripts in Bacillus thuringiensis and Escherichia coli. J Mol Biol. 1986 Sep 5;191(1):1–11. doi: 10.1016/0022-2836(86)90417-1. [DOI] [PubMed] [Google Scholar]
  36. Ward E. S., Ellar D. J. Cloning and expression of two homologous genes of Bacillus thuringiensis subsp. israelensis which encode 130-kilodalton mosquitocidal proteins. J Bacteriol. 1988 Feb;170(2):727–735. doi: 10.1128/jb.170.2.727-735.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ward E. S., Ellar D. J. Nucleotide sequence of a Bacillus thuringiensis var. israelensis gene encoding a 130 kDa delta-endotoxin. Nucleic Acids Res. 1987 Sep 11;15(17):7195–7195. doi: 10.1093/nar/15.17.7195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ward E. S., Ellar D. J., Todd J. A. Cloning and expression in Escherichia coli of the insecticidal delta-endotoxin gene of Bacillus thuringiensis var. israelensis. FEBS Lett. 1984 Oct 1;175(2):377–382. doi: 10.1016/0014-5793(84)80772-3. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES