Abstract
A newly engineered strain, denominated BIOT185, was constructed through integrating the cry8Ca2 gene into the endogenous plasmid of BT185 (contains cry8Ea1) by homologous recombination. The thermosensitive plasmid vector was eliminated by the rising temperature of recombinant cultures. No antibiotic gene or other unnecessary genes were introduced to the new strain. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis and Western blot analysis demonstrated that the cry8Ca2 gene was expressed normally and produced a 130-kDa protein in the BIOT185 strain. Bioassay results showed that the new strain had high toxicity to the pests Anomala corpulenta and Holotrichia parallela, which often damage the same fields.
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Acknowledgements
We are very grateful to Dr. Qinglei Wang for the supply of test larvae and for the help with the bioassays. This work was supported by Project 973 (No. 2009CB118902, 2007CB109203), Plan 863 (No.2006AA10A212), and 2009ZX08009-030B, 2009ZX08009-031B.
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Liu, J., Yan, G., Shu, C. et al. Construction of a Bacillus thuringiensis engineered strain with high toxicity and broad pesticidal spectrum against coleopteran insects. Appl Microbiol Biotechnol 87, 243–249 (2010). https://doi.org/10.1007/s00253-010-2479-5
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DOI: https://doi.org/10.1007/s00253-010-2479-5