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Integration of a Recombinant Chitinase into Bacillus thuringiensis Parasporal Insecticidal Crystal

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Abstract

Chitinases have been successfully used in combination with Bacillus thuringiensis delta-endotoxins forming crystals in order to enhance their insecticidal activities. In this context, we opted for promoting the chitinase integration into these crystals. Thus, we engineered, for the first time, a fusion protein (CDF) consisting of the chitinase Chi255 and the carboxy-terminal half of Cry1Ac, both from B. thuringiensis subsp. kurstaki. The constructed transcriptional fusion (chi255Δsp–CTcry1Ac) was cloned into a shuttle vector (Escherichia coli/B. thuringiensis) downstream the sporulation-dependent promoters BtI–BtII and upstream the cry1Ac transcription terminator. The resulting plasmid, named pF, was transferred by electroporation to crystalliferous B. thuringiensis strain BNS3 and acrystalliferous strain BNS3Cry−. The functionality of the chimerical chitinase was demonstrated by an improvement of the relative chitinolytic activity of the recombinant strain BNS3/pF by 2.5 folds. Western blot analyses showed that, despite of the instability of CDF when expressed in the acrystalliferous strain, the C-terminal half of Cry1Ac succeeded to allow the integration of the chimerical chitinase into the crystal of BNS3. The recombinant strain BNS3/pF (LC50 = 144.06 μg g−1) was 1.5 times more active against Ephestia kuehniella larvae than the wild strain (LC50 = 212.10 μg g−1).

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Acknowledgement

This work was supported by grants from the Tunisian «Ministère de l’Enseignement Supérieur et de la Recherche Scientifique (MESRS)».

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Authors and Affiliations

  1. Laboratory of Biopesticides, Centre of Biotechnology of Sfax, P.O. Box 1177, 3018, Sfax, Tunisia

    Fatma Driss, Souad Rouis, Hichem Azzouz, Slim Tounsi, Nabil Zouari & Samir Jaoua

  2. Department of Biological and Environmental Sciences, University of Qatar, College of Arts & Sciences, P.O. Box 2713, Doha, Qatar

    Samir Jaoua

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  1. Fatma Driss
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  2. Souad Rouis
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  3. Hichem Azzouz
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  4. Slim Tounsi
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  5. Nabil Zouari
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  6. Samir Jaoua
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Correspondence to Samir Jaoua.

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Driss, F., Rouis, S., Azzouz, H. et al. Integration of a Recombinant Chitinase into Bacillus thuringiensis Parasporal Insecticidal Crystal. Curr Microbiol 62, 281–288 (2011). https://doi.org/10.1007/s00284-010-9704-4

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  • DOI: https://doi.org/10.1007/s00284-010-9704-4

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