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Cry2A toxins from Bacillus thuringiensis expressed in insect cells are toxic to two lepidopteran insects

  • G. M. S. Lima
  • R. W. S. Aguiar
  • R. F. T. Corrêa
  • E. S. Martins
  • A. C. M. Gomes
  • T. Nagata
  • M. T. De-Souza
  • R. G. Monnerat
  • B. M. Ribeiro
Original Paper

Abstract

The cry2Aa and cry2Ab genes from a Brazilian Bacillus thuringiensis strain were introduced into the genome of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) in order to evaluate the heterologous proteins expression in insect cells and their toxicity to different insects. The recombinant viruses (vAcCry2Aa and vSynCry2Ab) were amplified in Trichoplusia ni (BTI-Tn5B1-4) cells and used to infect Spodoptera frugiperda larvae. Total extracts from S. frugiperda infected with the recombinant viruses were analysed by SDS-PAGE, which detected the presence of polypeptides around 65 kDa. Cuboid-shaped protein crystals were observed in insect extracts by light and scanning electron microscopy. Bioassays, using the heterologous proteins showed toxicity against second instar A. gemmatalis larvae (Cry2Aa) with a LC50 of 1.03 μg/ml and second instar S. frugiperda larvae (Cry2Ab) with a LC50 of 3.45 μg/ml. No toxic activity was detected for Aedes aegypti and Culex quinquenfaciatus.

Keywords

Baculovirus Cry2Aa Cry2Ab Bacillus thuringiensis Lepidopteran insects 

Notes

Acknowledgements

We are grateful to Embrapa Recursos Genéticos e Biotecnologia for supplying insect larvae. This work was supported by the following Brazilian agencies: PRONEX, FAPDF, CNPq, FINATEC and CAPES.

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • G. M. S. Lima
    • 1
  • R. W. S. Aguiar
    • 1
  • R. F. T. Corrêa
    • 1
  • E. S. Martins
    • 1
    • 2
  • A. C. M. Gomes
    • 2
  • T. Nagata
    • 1
  • M. T. De-Souza
    • 1
  • R. G. Monnerat
    • 2
  • B. M. Ribeiro
    • 1
  1. 1.Department of Cell BiologyUniversity of BrasíliaBrasiliaBrazil
  2. 2.Embrapa – Recursos Genéticos e BiotecnologiaBrasiliaBrazil

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