Dot Blot Enzyme-Linked Immunosorbent Assay for Monitoring the Fate of Insecticidal Toxins from Bacillus thuringiensis in Soil.

Applied and Environmental Microbiology
H Tapp, G Stotzky

Abstract

The release of transgenic plants and microorganisms expressing truncated genes from Bacillus thuringiensis that code for active insecticidal toxins rather than for the inactive protoxins could result in the accumulation of these active proteins in soil, especially when bound on clay minerals and other soil particles. To monitor the fate of these toxins in soil, a dot blot enzyme-linked immunosorbent assay (ELISA) that detects free and particle-bound toxins from B. thuringiensis subsp. kurstaki and subsp. tenebrionis was developed. The lower limit of detection of the toxins, either free or adsorbed or bound on the clay minerals montmorillonite (M) or kaolinite (K) or on the clay-particle-size fraction separated from soil (by sedimentation according to Stokes' Law), was approximately 3 ng. Antibodies (Ab) to the toxins from B. thuringiensis subsp. kurstaki and from B. thuringiensis subsp. thuringiensis were raised in goats and rabbits, respectively, and each Ab was rendered specific by adsorption onto CNBr-activated Sepharose coupled with the other toxin. The preadsorbed Ab were specific for the toxins from both subspecies, both free and bound on M, K, or the clay-particle-size fraction of soil. The toxins that were added to ster...Continue Reading

References

Jul 31, 1990·Biochemical and Biophysical Research Communications·S C MacIntoshR L Fuchs
Jun 1, 1989·Microbiological Reviews·H Höfte, H R Whiteley
May 1, 1989·Applied and Environmental Microbiology·Y S ZhuP Filner

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Citations

Jul 11, 2009·Organic & Biomolecular Chemistry·Sabrina LusvarghiBruce Alan Armitage
Aug 17, 2002·The Plant Journal : for Cell and Molecular Biology·Astrid T Groot, Marcel Dicke
Jan 22, 1998·Canadian Journal of Microbiology·H Tapp, G Stotzky
Jun 3, 2008·Journal of Agricultural and Food Chemistry·Haiyan WangJianmin Wu

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