Production of an insecticidal crystal protein from Bacillus thuringiensis by the methylotroph Methylobacterium extorquens

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DOIResolve DOI: http://doi.org/10.1128/AEM.00598-08
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TypeArticle
Journal titleApplied and Environmental Microbiology
Volume74
Issue16
Pages51785182; # of pages: 5
SubjectBacillus thuringiensis; bio; Methylobacterium extorquens; Protein
AbstractThe 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 δ-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.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada; NRC Biotechnology Research Institute
Peer reviewedNo
NRC number47842
NPARC number12390247
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Record identifier47721f9d-b1a2-4c29-8e41-00845a1991f0
Record created2009-10-26
Record modified2016-05-09
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