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AuthorDriss, Fatma
AuthorRouis, Souad
AuthorAzzouz, Hichem
AuthorTounsi, Slim
AuthorZouari, Nabil
AuthorJaoua, Samir
Available date2015-12-23T06:50:08Z
Publication Date2011-01
Publication NameCurrent Microbiology
CitationDriss, F., Rouis, S., Azzouz, H., et al, (2011). "Integration of a Recombinant Chitinase into Bacillus thuringiensis", Current Microbiology, 62(1), 281–288.
ISSN0343-8651
URIhttp://hdl.handle.net/10576/3976
URIhttp://dx.doi.org/10.1007/s00284-010-9704-4
AbstractChitinases 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 (LC(50) = 144.06 μg g(-1)) was 1.5 times more active against Ephestia kuehniella larvae than the wild strain (LC(50) = 212.10 μg g(-1)).
SponsorTunisian Ministere de l’Enseignement Superieur et de la Recherche Scientifique (MESRS)
Languageen
PublisherSpringer-Verlag
Subjectcry1Ac Gene
Gene Chitinase
Signal Peptide-encoding Sequence
Chitinolytic Activity
Recombinant Strain
TitleIntegration of a Recombinant Chitinase into Bacillus thuringiensis Parasporal Insecticidal Crystal
TypeArticle
Pagination281-288
Issue Number1
Volume Number62
ESSN1432-0991
dc.accessType Abstract Only


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