Improving Cry8Ka toxin activity towards the cotton boll weevil (Anthonomus grandis)

BMC Biotechnology

Gustavo R Oliveira³  Maria CM Silva⁶  Wagner A Lucena⁵  Erich YT Nakasu⁶  Alexandre AP Firmino⁵  Magda A Beneventi⁶  Djair SL Souza⁴  José E Gomes⁶  José DA de Souza⁶  Daniel J Rigden²  Hudson B Ramos⁴  Carlos R Soccol³  Maria F Grossi-de-Sa¹ 

[1] Pós-Graduação em Ciências Genômicas e Biotecnologia - UCB, Brasília, DF, Brasil

[2] Institute of Integrative Biology, University of Liverpool, Liverpool, UK

[3] Programa de Pós-Graduação em Processos Biotecnológicos-UFPR, Curitiba, PR, Brasil

[4] Departamento de Biologia Celular, Universidade de Brasília - UnB, Brasília, DF, Brasil

[5] Programa de Pós-graduação em Biologia Celular e Molecular, UFRGS, Porto Alegre, RS, Brasil

[6] Embrapa Recursos Genéticos e Biotecnologia, PqEB- Final W5 Norte -Brasília, DF, Brasil

关键词: Molecular modeling;    Phage display;    DNA shuffling;    Cotton;    Bacillus thuringiensis;    Anthonomus grandis;   

Others  :  1145824
DOI  :  10.1186/1472-6750-11-85

 received in 2011-04-05, accepted in 2011-09-09,  发布年份 2011

【 摘 要 】

Background

The cotton boll weevil (Anthonomus grandis) is a serious insect-pest in the Americas, particularly in Brazil. The use of chemical or biological insect control is not effective against the cotton boll weevil because of its endophytic life style. Therefore, the use of biotechnological tools to produce insect-resistant transgenic plants represents an important strategy to reduce the damage to cotton plants caused by the boll weevil. The present study focuses on the identification of novel molecules that show improved toxicity against the cotton boll weevil. In vitro directed molecular evolution through DNA shuffling and phage display screening was applied to enhance the insecticidal activity of variants of the Cry8Ka1 protein of Bacillus thuringiensis.

Results

Bioassays carried out with A. grandis larvae revealed that the LC₅₀ of the screened mutant Cry8Ka5 toxin was 3.15-fold higher than the wild-type Cry8Ka1 toxin. Homology modelling of Cry8Ka1 and the Cry8Ka5 mutant suggested that both proteins retained the typical three-domain Cry family structure. The mutated residues were located mostly in loops and appeared unlikely to interfere with molecular stability.

Conclusions

The improved toxicity of the Cry8Ka5 mutant obtained in this study will allow the generation of a transgenic cotton event with improved potential to control A. grandis.

【 授权许可】

   
2011 Oliveira et al; licensee BioMed Central Ltd.

【 图 表 】

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