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Constructing Bacillus thuringiensis strain that co-expresses Cry2Aa and chitinase

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Abstract

A triple recombineering technique was used with plasmid pHT315 to produce pHTEC, a construct carrying chitinase and cry2Aa genes from Bacillus thuringiensis subsp. kurstaki 4.0718. Transformation of wild-type B. thuringiensis strain HD73 and the acrystalliferous strain Cry-B with pHTEC resulted in the recovery of recombinant strains that expressed Cry2Aa as cubic crystals in the cell pellet and soluble chitinase protein. The toxicity of HD73 (pHTEC) against Helicoverpa armigera larvae increased sevenfold when compared with HD73 (pHT315) harboring pHT315 vector. The triple recombineering protocol was optimized by comparing recombination efficacy mediated by RecE/RecT and Redα/Redβ and by using single-strand DNA as substrate.

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Acknowledgments

This investigation was supported by National Natural Science Foundation of China (31200004), Specialized Research Fund for the Doctoral Program of Higher Education from the Ministry of Education of China (2011430620005), Hunan Provincial Natural Science Foundation of China (10JJ6042), Scientific Research Fund of Hunan Provincial Education Department (10K041).

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Authors and Affiliations

  1. Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, College of Life Science, Hunan Normal University, Changsha, China

    Shengbiao Hu, Xu Zhang, Yusheng Li, Xuezhi Ding, Xiaofeng Hu, Qi Yang & Liqiu Xia

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  1. Shengbiao Hu
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  2. Xu Zhang
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  3. Yusheng Li
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  4. Xuezhi Ding
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  5. Xiaofeng Hu
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  6. Qi Yang
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  7. Liqiu Xia
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Corresponding author

Correspondence to Liqiu Xia.

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Shengbiao Hu and Xu Zhang are Co-first authors.

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Hu, S., Zhang, X., Li, Y. et al. Constructing Bacillus thuringiensis strain that co-expresses Cry2Aa and chitinase. Biotechnol Lett 35, 1045–1051 (2013). https://doi.org/10.1007/s10529-013-1171-0

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  • DOI: https://doi.org/10.1007/s10529-013-1171-0

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