Construction of a Bacillus thuringiensis engineered strain with high toxicity and broad pesticidal spectrum against coleopteran insects |
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| Authors: | Jingjing Liu Guixin Yan Changlong Shu Can Zhao Chunqin Liu Fuping Song Lin Zhou Junlan Ma Jie Zhang Dafang Huang |
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| Institution: | (1) State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, People’s Republic of China;(2) Cangzhou Academy of Agricultural and Forestry Sciences, Cangzhou, 061001, People’s Republic of China;(3) Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China |
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| 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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