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PHYSIOLOGY AND BIOTECHNOLOGY

Optimization of Cry3A Yields in Bacillus thuringiensis by Use of Sporulation-Dependent Promoters in Combination with the STAB-SD mRNA Sequence

Hyun-Woo Park, Baoxue Ge, Leah S. Bauer, Brian A. Federici
Hyun-Woo Park
Department of Entomology and
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Baoxue Ge
Interdepartmental Graduate Program in Genetics, University of California, Riverside, California 92521, and
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Leah S. Bauer
USDA Forest Service, North Central Forest Experiment Station, Michigan State University, East Lansing, Michigan 48823
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Brian A. Federici
Department of Entomology and
Interdepartmental Graduate Program in Genetics, University of California, Riverside, California 92521, and
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DOI: 10.1128/AEM.64.10.3932-3938.1998
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ABSTRACT

The insecticidal activity of Bacillus thuringiensisstrains toxic to coleopterous insects is due to Cry3 proteins assembled into small rectangular crystals. Toxin synthesis in these strains is dependent primarily upon a promoter that is active in the stationary phase and a STAB-SD sequence that stabilizes the cry3transcript-ribosome complex. Here we show that significantly higher yields of Cry3A can be obtained by using dual sporulation-dependentcyt1Aa promoters to drive the expression ofcry3Aa when the STAB-SD sequence is included in the construct. The Cry3A yield per unit of culture medium obtained with this expression system was 12.7-fold greater than that produced by DSM 2803, the wild-type strain of B. thuringiensis from which Cry3Aa was originally described, and 1.4-fold greater than that produced by NB176, a mutant of the same strain containing two or three copies ofcry3Aa, which is the active ingredient of the commercial product Novodor, used for control of beetle pests. The toxicities of Cry3A produced with this construct or the wild-type strain were similar when assayed against larvae of the cottonwood leaf beetle,Chrysomela scripta. The volume of Cry3A crystals produced with cyt1Aa promoters and the STAB-SD sequence was 1.3-fold that of typical bipyramidal Cry1 crystals toxic to lepidopterous insects. The dual-promoter/STAB-SD system offers an additional method for potentially improving the efficacy of insecticides based onB. thuringiensis.

  • Copyright © 1998 American Society for Microbiology
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Optimization of Cry3A Yields in Bacillus thuringiensis by Use of Sporulation-Dependent Promoters in Combination with the STAB-SD mRNA Sequence
Hyun-Woo Park, Baoxue Ge, Leah S. Bauer, Brian A. Federici
Applied and Environmental Microbiology Oct 1998, 64 (10) 3932-3938; DOI: 10.1128/AEM.64.10.3932-3938.1998

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Optimization of Cry3A Yields in Bacillus thuringiensis by Use of Sporulation-Dependent Promoters in Combination with the STAB-SD mRNA Sequence
Hyun-Woo Park, Baoxue Ge, Leah S. Bauer, Brian A. Federici
Applied and Environmental Microbiology Oct 1998, 64 (10) 3932-3938; DOI: 10.1128/AEM.64.10.3932-3938.1998
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KEYWORDS

Bacillus thuringiensis
Bacterial Proteins
Endotoxins
Promoter Regions, Genetic

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