Display Settings:

Format

Send to:

Choose Destination
See comment in PubMed Commons below
Appl Environ Microbiol. 1998 Aug;64(8):2995-3003.

Phage display of a biologically active Bacillus thuringiensis toxin.

Author information

  • 1Department of Entomology, University of Georgia, Athens, Georgia 30602, USA.

Abstract

Activated forms of Bacillus thuringiensis insecticidal toxins have consistently been found to form insoluble and inactive precipitates when they are expressed in Escherichia coli. Genetic engineering of these proteins to improve their effectiveness as biological pesticides would be greatly facilitated by the ability to express them in E. coli, since the molecular biology tools available for Bacillus are limited. To this end, we show that activated B. thuringiensis toxin (Cry1Ac) can be expressed in E. coli as a translational fusion with the minor phage coat protein of filamentous phage. Phage particles displaying this fusion protein were viable, infectious, and as lethal as pure toxin on a molar basis when the phage particles were fed to insects susceptible to native Cry1Ac. Enzyme-linked immunosorbent assay and Western blot analysis showed the fusion protein to be antigenically equivalent to native toxin, and micropanning with anti-Cry1Ac antibody was positive for the toxin-expressing phage. Phage display of B. thuringiensis toxins has many advantages over previous expression systems for these proteins and should make it possible to construct large libraries of toxin variants for screening or biopanning.

PMID:
9687463
[PubMed - indexed for MEDLINE]
PMCID:
PMC106805
Free PMC Article

Images from this publication.See all images (4)Free text

FIG. 1
FIG. 2
FIG. 3
FIG. 4
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Icon for HighWire Icon for PubMed Central
    Loading ...
    Write to the Help Desk