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Engineered Cry1Ac-Cry9Aa hybrid Bacillus thuringiensis delta-endotoxin with improved insecticidal activity against Helicoverpa armigera

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Abstract

Recombinant Bt construct was prepared by exchange of pore forming domain I with cry1Ac to cry9Aa gene by overlap extension PCR (OE-PCR) technique. Construction of cry1Ac-cry9Aa was accomplished by six base pair homology at 3′ ends of PCR products of domain I of cry1Ac and domain II and III of cry9Aa. The recombinant toxin was also modified by deletion of N-terminal alpha helix-1 of recombinant toxin. Both Cry toxins were expressed in E. coli BL21(DE3) plysS and purified by His-tag purification. Upon insect bioassay analysis against devastating crop pest Helicoverpa armigera, toxicity of recombinant toxin was found around fivefold higher than native Cry1Ac while alpha helix-1 deleted N-terminal modified toxin did not resulted in significant increase in toxicity. The recombinant Cry toxins such as Cry1Ac-Cry9Aa and Cry1Ac-Cry9AaMod may be used for insect pest control.

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Fig. 1
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Abbreviations

Bt :

Bacillus thuringiensis

cry1Ac-cry9Aa :

Hybrid toxin with Cry1Ac domain I and Cry9Aa domain II and III

cry1Ac-cry9AaMod :

N-terminal alpha helix-1 deletion in cry1Ac-cry9Aa

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Acknowledgements

Jigar V. Shah would like to thank University Grant commission (UGC-RFMS), New Delhi for providing research fellowship.

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Authors and Affiliations

  1. Ground Floor Lab, Department of Microbiology and Biotechnology Centre, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, India

    Jigar V. Shah, Rakeshkumar Yadav & Sanjay S. Ingle

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  1. Jigar V. Shah
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  2. Rakeshkumar Yadav
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  3. Sanjay S. Ingle
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Correspondence to Sanjay S. Ingle.

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Communicated by Erko Stackebrandt.

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Shah, J.V., Yadav, R. & Ingle, S.S. Engineered Cry1Ac-Cry9Aa hybrid Bacillus thuringiensis delta-endotoxin with improved insecticidal activity against Helicoverpa armigera . Arch Microbiol 199, 1069–1075 (2017). https://doi.org/10.1007/s00203-017-1407-9

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  • DOI: https://doi.org/10.1007/s00203-017-1407-9

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