Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 87, Issue 1, Pages 243-249Publisher
SPRINGER
DOI: 10.1007/s00253-010-2479-5
Keywords
Bacillus thuringiensis; Engineered strain; Homologous recombination; Anomala corpulenta; Holotrichia parallela
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Funding
- Project 973 [2009CB118902, 2007CB109203]
- Plan 863 [2006AA10A212]
- [2009ZX08009-030B]
- [2009ZX08009-031B]
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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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