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. 1993 Apr;59(4):1131–1137. doi: 10.1128/aem.59.4.1131-1137.1993

Cloning of a novel cryIC-type gene from a strain of Bacillus thuringiensis subsp. galleriae.

S Kalman 1, K L Kiehne 1, J L Libs 1, T Yamamoto 1
PMCID: PMC202250  PMID: 8476286

Abstract

A novel cryIC-type gene was isolated from a strain of Bacillus thuringiensis subsp. galleriae. A new polymerase chain reaction (PCR) technique with a set of several oligonucleotide primer pairs specific to the cryIC gene was used to screen a number of B. thuringiensis strains. PCR amplified several DNA fragments ranging from 100 bp to 1 kb for B. thuringiensis strains containing a cryIC gene. PCR fragments amplified from the Bacillus thuringiensis subsp. galleriae HD29 DNA differed from the fragments amplified from other cryIC-containing strains, indicating strain HD29 contained a novel cryIC-type gene. To isolate crystal genes homologous to cryIC, an HD29 gene library was probed with a 984-bp fragment of the amino-terminal coding region of the cryIC gene cloned from Bacillus thuringiensis subsp. aizawai HD229. A putative toxin gene was isolated from a phage that hybridized strongly to the cryIC probe. Translation of the putative toxin DNA sequence revealed an open reading frame of 1,176 amino acids whose predicted molecular mass was 132.8 kDa. Comparisons of the toxin gene sequence with sequences of other cry genes indicated that this gene is a subclass of cryIC. We propose to designate this gene cryIC(b). In Escherichia coli, the cryIC(b) gene produced a protein of approximately 130 kDa toxic to Spodoptera exigua and Trichoplusia ni.

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Selected References

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  1. Bossé M., Masson L., Brousseau R. Nucleotide sequence of a novel crystal protein gene isolated from Bacillus thuringiensis subspecies kenyae. Nucleic Acids Res. 1990 Dec 25;18(24):7443–7443. doi: 10.1093/nar/18.24.7443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brown K. L., Whiteley H. R. Isolation of a Bacillus thuringiensis RNA polymerase capable of transcribing crystal protein genes. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4166–4170. doi: 10.1073/pnas.85.12.4166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown K. L., Whiteley H. R. Isolation of the second Bacillus thuringiensis RNA polymerase that transcribes from a crystal protein gene promoter. J Bacteriol. 1990 Dec;172(12):6682–6688. doi: 10.1128/jb.172.12.6682-6688.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carozzi N. B., Kramer V. C., Warren G. W., Evola S., Koziel M. G. Prediction of insecticidal activity of Bacillus thuringiensis strains by polymerase chain reaction product profiles. Appl Environ Microbiol. 1991 Nov;57(11):3057–3061. doi: 10.1128/aem.57.11.3057-3061.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chambers J. A., Jelen A., Gilbert M. P., Jany C. S., Johnson T. B., Gawron-Burke C. Isolation and characterization of a novel insecticidal crystal protein gene from Bacillus thuringiensis subsp. aizawai. J Bacteriol. 1991 Jul;173(13):3966–3976. doi: 10.1128/jb.173.13.3966-3976.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Honée G., Convents D., Van Rie J., Jansens S., Peferoen M., Visser B. The C-terminal domain of the toxic fragment of a Bacillus thuringiensis crystal protein determines receptor binding. Mol Microbiol. 1991 Nov;5(11):2799–2806. doi: 10.1111/j.1365-2958.1991.tb01988.x. [DOI] [PubMed] [Google Scholar]
  8. Honée G., van der Salm T., Visser B. Nucleotide sequence of crystal protein gene isolated from B. thuringiensis subspecies entomocidus 60.5 coding for a toxin highly active against Spodoptera species. Nucleic Acids Res. 1988 Jul 11;16(13):6240–6240. doi: 10.1093/nar/16.13.6240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Höfte H., Whiteley H. R. Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev. 1989 Jun;53(2):242–255. doi: 10.1128/mr.53.2.242-255.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Li J. D., Carroll J., Ellar D. J. Crystal structure of insecticidal delta-endotoxin from Bacillus thuringiensis at 2.5 A resolution. Nature. 1991 Oct 31;353(6347):815–821. doi: 10.1038/353815a0. [DOI] [PubMed] [Google Scholar]
  12. Moran C. P., Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982;186(3):339–346. doi: 10.1007/BF00729452. [DOI] [PubMed] [Google Scholar]
  13. Nicholls C. N., Ahmad W., Ellar D. J. Evidence for two different types of insecticidal P2 toxins with dual specificity in Bacillus thuringiensis subspecies. J Bacteriol. 1989 Sep;171(9):5141–5147. doi: 10.1128/jb.171.9.5141-5147.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sanchis V., Lereclus D., Menou G., Chaufaux J., Guo S., Lecadet M. M. Nucleotide sequence and analysis of the N-terminal coding region of the Spodoptera-active delta-endotoxin gene of Bacillus thuringiensis aizawai 7.29. Mol Microbiol. 1989 Feb;3(2):229–238. doi: 10.1111/j.1365-2958.1989.tb01812.x. [DOI] [PubMed] [Google Scholar]
  15. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Visser B. A screening for the presence of four different crystal protein gene types in 25 Bacillus thuringiensis strains. FEMS Microbiol Lett. 1989 Apr;49(2-3):121–124. doi: 10.1016/0378-1097(89)90024-4. [DOI] [PubMed] [Google Scholar]
  17. Visser B., Munsterman E., Stoker A., Dirkse W. G. A novel Bacillus thuringiensis gene encoding a Spodoptera exigua-specific crystal protein. J Bacteriol. 1990 Dec;172(12):6783–6788. doi: 10.1128/jb.172.12.6783-6788.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]

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