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Characterization of the cry1Ac17 Gene from an Indigenous Strain of Bacillus thuringiensis subsp. kenyae

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Abstract

An indigenously isolated strain of Bacillus thuringiensis subsp. kenyae exhibited toxicity against lepidopteran as well as dipteran insects. The lepidopteran active cry1Ac protoxin gene coding sequence of 3.5 kb from this strain was cloned into vector pET28a(+). However, it could not be expressed in commonly used Escherichia coli expression hosts, BL21(DE3) and BL21(DE3)pLysS. This gene is classified as cry1Ac17 in the B. thuringiensis toxic nomenclature database. The coding sequence of this gene revealed that it contains about 3% codons, which are not efficiently translated by these expression hosts. Hence, this gene was expressed in a modified expression host, Epicurian coli BL21-Codonplus (DE3)-RIL. The expression of gene yielded a 130-kDa Cry1Ac17 protein. The protein was purified and its toxicity was tested against economically important insect pests, viz., Helicoverpa armigera and Spodoptera litura. LC50 values obtained against these insects were 0.1 ng/cm3 and 1231 ng/cm2, respectively. The higher toxicity of Cry1Ac17 protein, compared to other Cry1Ac proteins, toward these pests demonstrates the potential of this isolate as an important candidate in the integrated resistance management program in India.

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Authors and Affiliations

  1. Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre (BARC), Trombay, Mumbai, 400 085, India

    Ramesh S. Hire, Tanaji K. Dongre & Stanislaus F. D’souza

  2. High Pressure Physics Division, Bhabha Atomic Research Centre (BARC), Trombay, Mumbai, 400 085, India

    Ravindra D. Makde

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  1. Ramesh S. Hire
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  2. Ravindra D. Makde
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  3. Tanaji K. Dongre
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  4. Stanislaus F. D’souza
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Correspondence to Tanaji K. Dongre.

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Hire, R.S., Makde, R.D., Dongre, T.K. et al. Characterization of the cry1Ac17 Gene from an Indigenous Strain of Bacillus thuringiensis subsp. kenyae . Curr Microbiol 57, 570–574 (2008). https://doi.org/10.1007/s00284-008-9244-3

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  • DOI: https://doi.org/10.1007/s00284-008-9244-3

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