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Appl Environ Microbiol. 1996 April; 62(4): 1369–1377.
PMCID: PMC167904

Identification of novel cry-type genes from Bacillus thuringiensis strains on the basis of restriction fragment length polymorphism of the PCR-amplified DNA.

Abstract

Two pairs of universal oligonucleotide primers were designed to probe the most conserved regions of all known cryI-type gene sequences so that the amplified PCR fragments of the DNA template from Bacillus thuringiensis strains may contain all possible cryI-type gene sequences. The restriction fragment length polymorphism (RFLP) patterns of the PCR-amplified fragments revealed that 14 distinct cry-type genes have been identified from 20 B. thuringiensis strains. Those cry-type genes included cryIA(a), cryIA(a), cryIA(b), cryIA(b), cryIA(c), cryIB, cryIC, cryIC, cryIC(b), cryID, cryIE, cryIF, cryIF, and cryIII (a dagger at the end of a gene designation indicates a novel cry-type gene determined by restriction mapping or DNA sequences). Among them, the sequences of cryIA(a), cryIA(b), cryIB, cryIC, cryIF, and cryIII were found to be different from the corresponding published cry gene sequences. Interestingly, five cry-type genes [cryIA(a)-, cryIB-, cryIC-, cryIC(b)-, and cryIF-type genes] and seven cry-type genes [cryIA(a)-, cryIA(b)-, cryIB-, cryIC-, cryIC(b)-, cryIF-, and cryIII-type genes] have been detected from B. thuringiensis subsp. morrisoni HD-12 and B. thuringiensis subsp. wuhanensis, respectively. Therefore, the PCR-RFLP typing system is a facile method to detect both known and novel cry genes existing in B. thuringiensis strains.

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Selected References

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  • Adang MJ, Staver MJ, Rocheleau TA, Leighton J, Barker RF, Thompson DV. 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. [PubMed]
  • Andrews RE, Jr, Faust RM, Wabiko H, Raymond KC, Bulla LA., Jr The biotechnology of Bacillus thuringiensis. Crit Rev Biotechnol. 1987;6(2):163–232. [PubMed]
  • Birnboim HC. A rapid alkaline extraction method for the isolation of plasmid DNA. Methods Enzymol. 1983;100:243–255. [PubMed]
  • Bourque SN, Valéro JR, Mercier J, Lavoie MC, Levesque RC. Multiplex polymerase chain reaction for detection and differentiation of the microbial insecticide Bacillus thuringiensis. Appl Environ Microbiol. 1993 Feb;59(2):523–527. [PMC free article] [PubMed]
  • Brizzard BL, Whiteley HR. Nucleotide sequence of an additional crystal protein gene cloned from Bacillus thuringiensis subsp. thuringiensis. Nucleic Acids Res. 1988 Mar 25;16(6):2723–2724. [PMC free article] [PubMed]
  • Brown KL, Whiteley HR. Molecular characterization of two novel crystal protein genes from Bacillus thuringiensis subsp. thompsoni. J Bacteriol. 1992 Jan;174(2):549–557. [PMC free article] [PubMed]
  • Bulla LA, Jr, Bechtel DB, Kramer KJ, Shethna YI, Aronson AI, Fitz-James PC. Ultrastructure, physiology, and biochemistry of Bacillus thuringiensis. Crit Rev Microbiol. 1980;8(2):147–204. [PubMed]
  • Carozzi NB, Kramer VC, Warren GW, Evola S, Koziel MG. Prediction of insecticidal activity of Bacillus thuringiensis strains by polymerase chain reaction product profiles. Appl Environ Microbiol. 1991 Nov;57(11):3057–3061. [PMC free article] [PubMed]
  • Ceron J, Covarrubias L, Quintero R, Ortiz A, Ortiz M, Aranda E, Lina L, Bravo A. PCR analysis of the cryI insecticidal crystal family genes from Bacillus thuringiensis. Appl Environ Microbiol. 1994 Jan;60(1):353–356. [PMC free article] [PubMed]
  • Chak KF, Chao DC, Tseng MY, Kao SS, Tuan SJ, Feng TY. Determination and Distribution of cry-Type Genes of Bacillus thuringiensis Isolates from Taiwan. Appl Environ Microbiol. 1994 Jul;60(7):2415–2420. [PMC free article] [PubMed]
  • Chak KF, Young YM. Characterization of the Bacillus thuringiensis strains isolated from Taiwan. Proc Natl Sci Counc Repub China B. 1990 Jul;14(3):175–182. [PubMed]
  • Chambers JA, Jelen A, Gilbert MP, Jany CS, Johnson TB, 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. [PMC free article] [PubMed]
  • Chungjatupornchai W, Höfte H, Seurinck J, Angsuthanasombat C, Vaeck M. Common features of Bacillus thuringiensis toxins specific for Diptera and Lepidoptera. Eur J Biochem. 1988 Apr 5;173(1):9–16. [PubMed]
  • Dardenne F, Seurinck J, Lambert B, Peferoen M. Nucleotide sequence and deduced amino acid sequence of a cryIA(c) gene variant from Bacillus thuringiensis. Nucleic Acids Res. 1990 Sep 25;18(18):5546–5546. [PMC free article] [PubMed]
  • Donovan WP, Rupar MJ, Slaney AC, Malvar T, Gawron-Burke MC, Johnson TB. Characterization of two genes encoding Bacillus thuringiensis insecticidal crystal proteins toxic to Coleoptera species. Appl Environ Microbiol. 1992 Dec;58(12):3921–3927. [PMC free article] [PubMed]
  • González JM, Jr, Carlton BC. Patterns of plasmid DNA in crystalliferous and acrystalliferous strains of Bacillus thuringiensis. Plasmid. 1980 Jan;3(1):92–98. [PubMed]
  • Granum PE, Pinnavaia SM, Ellar DJ. Comparison of the in vivo and in vitro activity of the delta-endotoxin of Bacillus thuringiensis var. morrisoni (HD-12) and two of its constituent proteins after cloning and expression in Escherichia coli. Eur J Biochem. 1988 Mar 15;172(3):731–738. [PubMed]
  • Herrnstadt C, Gilroy TE, Sobieski DA, Bennett BD, Gaertner FH. Nucleotide sequence and deduced amino acid sequence of a coleopteran-active delta-endotoxin gene from Bacillus thuringiensis subsp. san diego. Gene. 1987;57(1):37–46. [PubMed]
  • Höfte H, Whiteley HR. Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev. 1989 Jun;53(2):242–255. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Kalman S, Kiehne KL, Libs JL, Yamamoto T. Cloning of a novel cryIC-type gene from a strain of Bacillus thuringiensis subsp. galleriae. Appl Environ Microbiol. 1993 Apr;59(4):1131–1137. [PMC free article] [PubMed]
  • Lambert B, Höfte H, Annys K, Jansens S, Soetaert P, Peferoen M. Novel Bacillus thuringiensis insecticidal crystal protein with a silent activity against coleopteran larvae. Appl Environ Microbiol. 1992 Aug;58(8):2536–2542. [PMC free article] [PubMed]
  • Martin PA, Travers RS. Worldwide Abundance and Distribution of Bacillus thuringiensis Isolates. Appl Environ Microbiol. 1989 Oct;55(10):2437–2442. [PMC free article] [PubMed]
  • Schnepf HE, Wong HC, Whiteley HR. The amino acid sequence of a crystal protein from Bacillus thuringiensis deduced from the DNA base sequence. J Biol Chem. 1985 May 25;260(10):6264–6272. [PubMed]
  • Sick A, Gaertner F, Wong A. Nucleotide sequence of a coleopteran-active toxin gene from a new isolate of Bacillus thuringiensis subsp. tolworthi. Nucleic Acids Res. 1990 Mar 11;18(5):1305–1305. [PMC free article] [PubMed]
  • Smulevitch SV, Osterman AL, Shevelev AB, Kaluger SV, Karasin AI, Kadyrov RM, Zagnitko OP, Chestukhina GG, Stepanov VM. Nucleotide sequence of a novel delta-endotoxin gene cryIg of Bacillus thuringiensis ssp. galleriae. FEBS Lett. 1991 Nov 18;293(1-2):25–28. [PubMed]
  • Tailor R, Tippett J, Gibb G, Pells S, Pike D, Jordan L, Ely S. Identification and characterization of a novel Bacillus thuringiensis delta-endotoxin entomocidal to coleopteran and lepidopteran larvae. Mol Microbiol. 1992 May;6(9):1211–1217. [PubMed]
  • Thorne L, Garduno F, Thompson T, Decker D, Zounes M, Wild M, Walfield AM, Pollock TJ. 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. [PMC free article] [PubMed]
  • Visser B, Munsterman E, Stoker A, Dirkse WG. A novel Bacillus thuringiensis gene encoding a Spodoptera exigua-specific crystal protein. J Bacteriol. 1990 Dec;172(12):6783–6788. [PMC free article] [PubMed]
  • Wabiko H, Raymond KC, Bulla LA., Jr Bacillus thuringiensis entomocidal protoxin gene sequence and gene product analysis. DNA. 1986 Aug;5(4):305–314. [PubMed]
  • Ward ES, Ellar DJ. 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. [PMC free article] [PubMed]

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