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Cloning, expression and activity of cry1Ia gene from Bacillus thuringiensis isolate

Published online by Cambridge University Press:  27 June 2008

Dou Li-Ming ,
Han Lan-Lan ,
Zhang Jie ,
He Kang-lai ,
Zhao Kui-Jun ,
Huang Da-Fang  and
Song Fu-Ping
Dou Li-Ming
Affiliation:
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China College of Agriculture, Northeast Agricultural University, Harbin 150030, China
Han Lan-Lan
Affiliation:
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China College of Agriculture, Northeast Agricultural University, Harbin 150030, China
Zhang Jie
Affiliation:
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China
He Kang-lai
Affiliation:
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China
Zhao Kui-Jun
Affiliation:
College of Agriculture, Northeast Agricultural University, Harbin 150030, China
Huang Da-Fang
Affiliation:
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Song Fu-Ping*
Affiliation:
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China
*
*Corresponding author. E-mail: [email protected]
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Abstract

A full-length cry1Ia gene fragment, obtained by polymerase chain reaction (PCR) amplification with a pair of primers designed according to cry1Ia-type gene sequences and DNA from Bacillus thuringiensis as template, was introduced into the expression vector pET-21b and transformed into Escherichia coli BL21(DE3). The molecular weight of the induced expression product was 81.2 kDa. The amino acid sequence of Cry1Ia was very different from sequences of 12 known Cry1Ia-type proteins. This gene was designated as cry1Ia8 by the International B. thuringiensis Insecticidal Protein Nomenclature Committee (accession number AF373207). The bioassay results indicated that Cry1Ia toxin protein showed distinct insecticidal activity against Ostrinia furnacalis and Plutella xylostella with LC50 of 0.268 μg/g and 2.227 μg/ml, respectively. It also had insecticidal activity against Leguminivora glycinivorella, but not against Pyrrhalta aenescens. The novel cry1Ia8 gene will be an important resource in constructing genetically engineered bacteria and transgenic plants for biocontrol of insect pests. It is also available for screening gene stacks to delay pest resistance.

Keywords

Bacillus thuringiensiscry1Ia genecloningexpressioninsecticidal activity
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 5 , Issue 1 , April 2008 , pp. 49 - 53
Copyright
Copyright © China Agricultural University 2008

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Footnotes

First published in Journal of Agricultural Biotechnology 2007, 15(6): 1053–1057

References

Angamuthu, S, Vanga, SR, Kumar, PA, Tewari, KK and Bhatnagar, RK (1998) Transformation of Nicotiana tabacum with a native cry1Ia5 gene confers complete protection against Heliothis armigera. Molecular Breeding 4: 473–478.Google Scholar
Choi, SK, Shin, BS, Kong, EM, Rho, HM and Park, SH (2000) Cloning of a new Bacillus thuringiensis cry1I-type crystal protein gene. Current Microbiology 41: 65–69.CrossRefGoogle ScholarPubMed
Crickmore, N, Zeigler, DR, Feitelson, J, et al. (1998) Revision of the nomenclature for the Bacillus thuringiensis pesticidal crystal proteins. Microbiology and Molecular Biology Reviews 62: 807.CrossRefGoogle ScholarPubMed
Diego, HS, Jorge, S, Alejandra, B and Graciela, B (2007) Toxicity of Bacillus thuringiensis δ-endotoxins against bean shoot borer (Epinotia aporema Wals.) larvae, a major soybean pest in Argentina. Journal of Invertebrate Pathology 94: 125–129.Google Scholar
Escudero, IR, Estela, A, Porcar, M, et al. (2006) Molecular and insecticidal characterization of a Cry1I protein toxic to insects of the families Noctuidae, Tortricidae, Plutellidae, and Chrysomelidae. Applied and Environmental Microbiology 72(7): 4796–4804.CrossRefGoogle Scholar
Kostichka, K, Warren, GW, Mullins, M, et al. (1996) Cloning of a cryV-type insecticidal protein gene from Bacillus thuringiensis: The cryV-encoded protein is expressed early in stationary phase. Journal of Bacteriology 178: 2141–2144.CrossRefGoogle ScholarPubMed
Liu, JH, Xue, J, Song, FP, Tan, JX, Zhang, J and Huang, DF (2006) Identification, cloning and activity of the insecticidal crystal protein genes from Bacillus thuringiensis isolate BtC008. Plant Protection 32(6): 22–26 (in Chinese with English abstract).Google Scholar
Ren, YB, Li, GX, Wang, QY, Huang, DF, Song, FP and Zhang, J (2004) Cloning and expression of cry1Aal4 gene from Bacillus thuringiensis. Journal of Agricultural Biotechnology 12(2): 188–191 (in Chinese with English abstract).Google Scholar
Russell, RM, Robertson, JL and Savin, NE (1977) POLO: a new computer program for probit analysis. Bulletin of the Entomological Society of America 23: 209–213.CrossRefGoogle Scholar
Sambrook, J and Russell, DW (2001) Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press.Google Scholar
Schnepf, E and Whiteley, R (1981) Cloning and expression of the Bacillus thuringiensis crystal protein gene in Escherichia coli. Proceedings of the National Academy of Sciences of the USA 78: 2893–2897.CrossRefGoogle ScholarPubMed
Sekar, V, Held, B, Tippett, J, et al. (1997) Biochemical and molecular characterization of the insecticidal fragment of CryV. Applied and Environmental Microbiology 63(7): 2798–2801.CrossRefGoogle ScholarPubMed
Shin, BS, Park, SH, Choi, SK, Koo, BT, Lee, ST and Kim, JI (1995) Distribution of cryV-type insecticidal protein genes in Bacillus thuringiensis and cloning of cryV-type genes from Bacillus thuringiensis subsp. kurstaki and Bacillus thuringiensis subsp. entomocidus. Applied and Environmental Microbiology 61(6): 2402–2407.CrossRefGoogle ScholarPubMed
Song, FP, Zhang, J, Ding, ZQ and Huang, DF (1998) Establishment of PCR-RFLP identification system of cry-type genes from Bacillus thuringiensis. Scientia Agricultura Sinica 31: 13–18 (in Chinese with English abstract).Google Scholar
Song, FP, Zhang, J, Gu, AX, et al. (2002) Analysis of the silent gene cry1I from Bacillus thuringiensis. Progress in Natural Science 12: 34–938 (in Chinese with English abstract).Google Scholar
Song, FP, Zhang, J, Gu, AX, et al. (2003) Identification of cry1I-type genes from Bacillus thuringiensis strains and characterization of a novel cry1I-type gene. Applied and Environmental Microbiology 69(9): 5207–5211.CrossRefGoogle ScholarPubMed
Tailor, R, Tippett, J, Gibb, G, et al. (1992) Identification and characterization of a novel Bacillus thuringiensis delta-endotoxin entomocidal to coleopteran and lepidopteran larvae. Molecular Microbiology 6: 1211–1217.CrossRefGoogle ScholarPubMed
Tounsi, S, Zouari, N and Jaoua, S (2003) Cloning and study of the expression of a novel cry1Ia-type gene from Bacillus thuringiensis subsp kurstaki. Journal of Applied Microbiology 95: 23–28.CrossRefGoogle ScholarPubMed
Zhang, J, Song, FP, Li, CY and Huang, DF (2002) Cloning and expression of cry3Aa7 gene from Bacillus thuringiensis strain toxic to coleopteran pests. Scientia Agricultura Sinica 35(6): 650–653 (in Chinese with English abstract).Google Scholar