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Characterisation and expression of a novel haplotype cry2A-type gene from Bacillus thuringiensis strain JF19-2

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Abstract

The Bacillus thuringiensis isolate JF19-2, a native organism found in soil samples from the Sichuan Basin in western China, contains a bipyramidal crystal harbouring one insecticidal crystal protein (about 70 kDa). Interestingly, a novel cry2A-type gene, encoding a 635 amino acid protein with 92% homology to cry2Ab1, was found and cloned from this strain. According to the nomenclature principles adopted for B. thuringiensis insecticidal crystal proteins, this gene represents a novel haplotype cry gene and was designated as cry2Ag1 by the Bacillus thuringiensis Pesticide Crystal Protein Nomenclature Committee. Cry2Ag1 was expressed in Escherichia coli BL21(DE3)pLysS cells, and the results of insecticidal activity assays showed that Cry2Ag1 was toxic to both Dipteran (Aedes aegypti) and Lepidopteran (Plutella xylostella and Helicoverpa armigera) pests. These results strongly suggest that the cloning of the cry2Ag1 gene is not only of interest to researchers studying insecticidal crystal genes, but may ultimately serve as a solution to the increasing resistance of pests to currently used insecticides.

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References

  • Alcantara EP, Aguda RM, Curtiss A, Dean DH, Cohen MB (2004) Bacillus thuringiensis δ-endotoxin binding to brush border membrane vesicles of rice stem borers. Arch Insect Biochem Physiol 55:169–177

    Article  CAS  PubMed  Google Scholar 

  • Boonserm P, Davis P, Ellar DJ, Li J (2005) Crystal structure of the mosquito-larvicidal toxin Cry4Ba and its biological implications. J Mol Biol 348:363–382

    Article  CAS  PubMed  Google Scholar 

  • Chen H, Tang W, Xu CG, Li XH, Lin YJ, Zhang QF (2005) Transgenic indica rice plants harboring a synthetic cry2A* gene of Bacillus thuringiensis exhibit enhanced resistance against lepidopteran rice pests. Theor Appl Genet 111:1330–1337

    Article  CAS  PubMed  Google Scholar 

  • Crickmore N, Zeigler DR, Feitelson J, Schnepf E, Van Rie J, Lereclus D, Baum J, Dean DH (1998) Revision of the nomenclature for the Bacillus thuringiensis pesticidal crystal proteins. Microbiol Mol Biol Rev 62:807–813

    CAS  PubMed  Google Scholar 

  • Galitsky N, Cody V, Wojtczak A, Ghosh D, Luft JR, Pangborn W, English L (2001) Structure of the insecticidal bacterial δ-endotoxin Cry3Bb1 of Bacillus thuringiensis. Acta Crystallogr D Biol Crystallogr 57:1099–1101

    Article  Google Scholar 

  • Grochulski P, Masson L, Borisova S, Pusztai-Carey M, Schwartz J, Brousseau R, Cygler M (1995) Bacillus thuringiensis CryIA(a) insecticidal toxin: crystal structure and channel formation. J Mol Biol 254:447–464

    Article  CAS  PubMed  Google Scholar 

  • Karim S, Dean DH (2000) Toxicity and receptor binding properties of Bacillus thuringiensis δ-endotoxins to the midgut brush border membrane vesicles of the rice leaf folders, Cnaphalocrocis medinalis and Marasmia patnalis. Curr Microbiol 41:276–283

    Article  CAS  PubMed  Google Scholar 

  • Lenin K, Mariam MA, Udayasuriyan V (2001) Expression of a cry2Aa gene in an acrystalliferous Bacillus thuringiensis strain and toxicity of Cry2Aa against Helicoverpa armigera. World J Microbiol Biotechnol 17:273–278

    Article  CAS  Google Scholar 

  • Li JD, Carroll J, Ellar DJ (1991) Crystal structure of insecticidal δ-endotoxin from Bacillus thuringiensis at 2.5 Å resolution. Nature 353:815–821

    Article  CAS  PubMed  Google Scholar 

  • Liang Y, Dean DH (1994) Location of a lepidopteran specificity region in insecticidal crystal protein CryIIA from Bacillus thuringiensis. Mol Microbiol 13(4):569–575

    Article  CAS  PubMed  Google Scholar 

  • McGaughey WH (1985) Insect resistance to the biological insecticide Bacillus thuringiensis. Science 229:193–195

    Article  PubMed  Google Scholar 

  • Morse RJ, Yamamoto T, Stroud RM (2001) Structure of Cry2Aa suggests an unexpected receptor binding epitope. Structure 9:409–417

    Article  CAS  PubMed  Google Scholar 

  • Romeis J, Meissle M, Bigler F (2006) Transgenic crops expressing Bacillus thuringiensis toxins and biological control. Nat Biotechnol 24(1):63–71

    Article  CAS  PubMed  Google Scholar 

  • Sambrook JE, Fritsch EF, Maaiatis T (2002) A laboratory manual of molecular cloning, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  • Sauka DH, Cozzi JG, Benintende GB (2005) Screening of cry2 genes in Bacillus thuringiensis isolates from Argentina. Antonie van Leeuwenhoek 88:163–165

    Article  CAS  PubMed  Google Scholar 

  • Schnepf E, Crickmore N, Rie JV, Lereclus D, Baum J, Feitelson J, Zeigler DR, Dean DH (1998) Bacillus thuringiensis and its pesticidal crystal proteins. Microbiol Mol Biol Rev 62:775–806

    CAS  PubMed  Google Scholar 

  • Servant F, Bru C, Carrere S, Courcelle E, Gouzy J, Peyruc D, Kahn D (2002) ProDom: Automated clustering of homologous domains. Brief Bioinform 3:246–251

    Article  CAS  PubMed  Google Scholar 

  • Sims SR (1997) Host activity spectrum of the CryIIA Bacillus thuringiensis subsp. kurstaki protein: effects on Lepidoptera, Diptera, and non-target arthropods. Southwest Entomol 22:139–404

    Google Scholar 

  • Zhu J, Tan FR, Tang J, Li YY, Zheng AP, Li P (2009) Characterization of insecticidal crystal protein cry gene of Bacillus thuringiensis from soil of Sichuan Basin, China and cloning of novel haplotypes cry gene. Ann Microbiol 59(1):1–8

    Article  CAS  Google Scholar 

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Acknowledgements

This study was supported by the Ministry of Agriculture of China for transgenic research (No. 2008ZX08009-003), National Basic Research Program of China (No. 2006CB101700), and Excellent Doctoral Dissertation Fund of Sichuan Agricultural University.

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Correspondence to Ping Li.

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Jun Zhu and Aiping Zheng contributed equally to this work.

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Zheng, A., Zhu, J., Tan, F. et al. Characterisation and expression of a novel haplotype cry2A-type gene from Bacillus thuringiensis strain JF19-2. Ann Microbiol 60, 129–134 (2010). https://doi.org/10.1007/s13213-009-0011-x

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  • DOI: https://doi.org/10.1007/s13213-009-0011-x

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