AEM
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Other Versions of this Article:
AEM.00598-08v1
74/16/5178    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Google Scholar
Right arrow Articles by Choi, Y. J.
Right arrow Articles by Míguez, C. B.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Choi, Y. J.
Right arrow Articles by Míguez, C. B.
Agricola
Right arrow Articles by Choi, Y. J.
Right arrow Articles by Míguez, C. B.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, August 2008, p. 5178-5182, Vol. 74, No. 16
0099-2240/08/$08.00+0     doi:10.1128/AEM.00598-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Production of an Insecticidal Crystal Protein from Bacillus thuringiensis by the Methylotroph Methylobacterium extorquens{triangledown}

Young J. Choi,1 J. Lawrence Gringorten,2 Louise Bélanger,1 Lyne Morel,1 Denis Bourque,1 Luke Masson,1,3 Denis Groleau,1 and Carlos B. Míguez1*

Microbial and Enzymatic Technology Group, Bioprocess Sector,1 Environmental Sector, Biotechnology Research Institute, National Research Council of Canada, 6100 Royalmount Avenue, Montreal, Quebec H4P 2R2, Canada,3 Canadian Forest Service, Great Lakes Forestry Centre, 1219 Queen Street E., Sault Ste. Marie, Ontario P6A 2E5, Canada2

Received 12 March 2008/ Accepted 9 June 2008

The Cry1Aa protein from Bacillus thuringiensis is an insecticidal protein that is highly active against several species of Lepidoptera. We cloned and expressed the cry1Aa gene in a plant-colonizing methylotroph, Methylobacterium extorquens, under the control of the strong M. extorquens AM1 methanol dehydrogenase promoter, PmxaF. Transmission electron microscopy revealed characteristic bipyramidal intracellular {delta}-endotoxin crystals similar to the crystalline inclusions formed by B. thuringiensis. Both the protoxin protein and the activated toxin were visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western analysis. In single-dose assays of the recombinant against the silkworm, Bombyx mori, both whole cells and cell lysates caused rapid feeding inhibition followed by mortality. The biomass and growth rate of recombinant cells in shake flask culture were similar to those of the wild-type strain, indicating a lack of fitness cost to the recombinant under controlled culture conditions. Recombinant Cry1Aa was expressed at a level of 4.5% of total M. extorquens cell protein. The potential benefits of modifying M. extorquens to deliver insecticidal Cry proteins for crop and forest protection are discussed.

* Corresponding author. Mailing address: Microbial and Enzymatic Technology Group, Bioprocess Sector, Biotechnology Research Institute, National Research Council of Canada, 6100 Royalmount Ave., Montreal, Quebec H4P 2R2, Canada. Phone: (514) 496-6280. Fax: (514) 496-7251. E-mail: carlos.miguez{at}nrc-cnrc.gc.ca

{triangledown} Published ahead of print on 13 June 2008.

Applied and Environmental Microbiology, August 2008, p. 5178-5182, Vol. 74, No. 16
0099-2240/08/$08.00+0     doi:10.1128/AEM.00598-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
J. Bacteriol. Microbiol. Mol. Biol. Rev. Eukaryot. Cell All ASM Journals

Copyright © 2008 by the American Society for Microbiology. All rights reserved.