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Efeitos da interação e toxicidade das proteínas Cry1 e Vip3Aa de Bacillus thuringiensis, Berliner em Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae)

  • Pavani, Claudio Damasceno
Publication Date
Jul 29, 2013
Repositório Institucional UNESP
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Bacillus thuringiensis is kwon as one of the best options for biological control of pests due to the entomophotogenic and specific action of some of its proteins. The Vip3 proteins, that are secreted during the vegetative growth of B. thuringiensis and Cry1, which are produced during the sporulation phase, act controlling important lepidopteran pests. One of the most important environmental risks associated to the use of insecticides is the potential for the development of increasing degrees of resistance by the target pests. The use of different entomopathogenic proteins produced by B. thuringiensis, as well as the studies involving their interactions might help overcoming the development of resistant pests. The proteins used in this study were prepared from recombinant Escherichia coli strains expressing a sole toxin. For the expression of the Cry1 proteins, the recombinant cells were inoculated in TB broth containing ampicillin and incubated at 28ºC during 60 h. While for the expression of the Vip3Aa protein the culture was made using LB 2X broth also containing ampicillin and induced with IPTG during incubation for 12 h at 190 rpm kept at 37ºC. The proteins were activated using 10% commercial trypsin, and observed on 12% SDS-PAGE gels. The bioassays were carried out using artificial semi-solid diet added to polystyrene trays with 128 wells measuring 1 cm of diameter. Fifty microliters of activated toxins were spread over the surface of the artificial diet contained wells and for each protein concentration, four repetitions were reproduced. Sixteen Spodoptera frugiperda first instar larvae were used adding to a total of 64 larvae for each protein concentration...

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