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Applied Microbiology and Biotechnology

, Volume 63, Issue 3, pp 300–306 | Cite as

A δ-endotoxin encoded in Pseudomonas fluorescens displays a high degree of insecticidal activity

  • R. Peng
  • A. Xiong
  • X. Li
  • H. Fuan
  • Q. YaoEmail author
Original Paper

Abstract

The short field-life of Bacillus thuringiensis (Bt) insecticidal crystal protein has limited its use. When the Bt toxin is produced in Pseudomonas fluorescens it can be encapsulated and retain its effectiveness for two to three times longer than other Bt formulations. In order to improve Bt expression, we have synthesized cryIA(c) Bt δ-endotoxin encoding region (GenBank AF537267) according to the usage codon of P. fluorescens and transformed the Bt toxin expression cassette into P. fluorescens strains. T7 RNA polymerase and the T7 promoter system were used to control expression of Bt toxin. SDS-PAGE and Western blotting assay revealed that the δ-endotoxin was expressed as 8% of the total protein in P. fluorescens. In in vitro tests, release of toxin from dead bacteria was demonstrated. Supplementation of diets with Bt toxin-containing Pseudomonas bacterium resulted in high mortality of cabbage butterfly (Pieris brassicae) larvae.

Keywords

Usage Codon Pseudomonas Fluorescens Insecticidal Activity Wild Type Gene Codon Usage Pattern 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

We are grateful to Reinhard Simon for providing E. coli S-17 and mini-Tn5 Tc, Tohru Nishihara for vector pNIN-215 and Kovach for pUT56.

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  1. 1.Agro-Biotech Research CenterShanghai Academy of Agricultural ScienceShanghaiChina

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