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The 20-kDa Protein of Bacillus thuringiensis subsp. israelensis Enhances Bacillus sphaericus 2362 Bin Toxin Synthesis

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Abstract

Improving the amount of protein endotoxins synthesized by Bacillus thuringiensis and B. sphaericus per unit of culture medium is important because higher yields typically correlate with higher insecticidal activity per unit weight of spore/toxin mixtures. Higher levels of synthesis can also result in larger crystals that could persist for longer periods in the environment. Improving endotoxin production in B. thuringiensis can be achieved by manipulating genetic elements that regulate protein synthesis at the transcriptional, translational, and even posttranslational levels. In the present study, we used a combination of genetic elements to improve yields of B. sphaericus 2362 binary toxin (Bin) in B. thuringiensis. Our results show that a 20-kDa chaperone-like protein, which occurs as the third open-reading frame in the cry11Aa operon, improves Bin yields when expression of the genes encoding this binary toxin is driven by the native bin promoter, cyt1A promoters, or a novel cyt1A-p/STAB-SD expression system, the latter of which yields maximal levels of Bin synthesis. The 20-kDa helper protein increased Bin toxin levels in B. thuringiensis by as much as 53% and concomitant toxicity by at least 90% when Bin was produced using the cyt1A promoters.

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Acknowledgments

We thank Jeffrey J. Johnson for technical assistance during this study. This research was supported in part by a grant (to B.A.F.) from the United States National Institutes of Health (AI45817).

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Authors and Affiliations

  1. John A. Mulrennan, Sr., Public Health Entomology Research and Education Center, College of Engineering Sciences, Technology, and Agriculture, Florida A & M University, 4000 Frankford Avenue, Panama City, FL, 32405, USA

    Hyun-Woo Park

  2. Department of Entomology, University of California, Riverside, Riverside, CA, 92521, USA

    Dennis K. Bideshi & Brian A. Federici

  3. Interdepartmental Graduate Programs in Genetics, Genomics, and Bioinformatics, and Cell, Molecular, and Developmental Biology, University of California, Riverside, Riverside, CA, 92521, USA

    Brian A. Federici

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  1. Hyun-Woo Park
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  2. Dennis K. Bideshi
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  3. Brian A. Federici
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Correspondence to Hyun-Woo Park.

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Park, HW., Bideshi, D.K. & Federici, B.A. The 20-kDa Protein of Bacillus thuringiensis subsp. israelensis Enhances Bacillus sphaericus 2362 Bin Toxin Synthesis. Curr Microbiol 55, 119–124 (2007). https://doi.org/10.1007/s00284-006-0359-0

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