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Construction of an environmental safe Bacillus thuringiensis engineered strain against Coleoptera

  • Applied genetics and molecular biotechnology
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Abstract

Cloning of new toxic genes from Bacillus thuringiensis (Bt) and construction of Bt engineered strains are two key strategies for bio-control of coleopteran pests in agriculture and forestry. In this study, we cloned a new cry3Aa-type gene, cry3Aa8, from wild Bt strain YC-03 against coleopteran, and constructed a Bt engineered strain, ACE-38, containing insecticidal protein-encoding gene cry3Aa8. The engineered strain, with almost four times of Cry3Aa yield compared with strain YC-03, was an antibiotic marker-free strain. Though no selective pressure was presented in the medium, cry3Aa8 in the engineered strain ACE-38 remained stable. The yield of Cry3Aa by strain ACE-38 reached 2.09 mg/ml in the optimized fermentation medium. The activity of strain ACE-38 against Plagiodera versicolora was tested, and the LC50 of ACE-38 cultures in the optimized fermentation medium was 1.13 μl/ml. Strain ACE-38 is a non-antibiotic Bt engineered strain with high Chrysomelidae toxicity and exhibits good fermentation property. The modified indigenous site-specific recombination system constructed in this study might be useful for the construction of Bt engineered strains containing genes that cannot be expressed in the indigenous site-specific recombination system using plasmid pBMB1205R.

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Acknowledgments

We acknowledged Dr. Sun Ming at Huazhong Agricultural University in China for providing vectors pHT304, pBMB1205R, pBMB1200 and Bacillus thuringiensis acrystalliferous mutant strain BMB171.

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Authors and Affiliations

  1. Key Laboratory of Agricultural and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, People’s Republic of China

    Yajun Yu, Yihui Yuan & Meiying Gao

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  1. Yajun Yu
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  2. Yihui Yuan
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  3. Meiying Gao
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Correspondence to Meiying Gao.

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Funding

This study was funded by the National Nature Science Foundation of China (No. 31170123) and the project of Chinese Academy of Sciences (KSZD-EW-Z-021-2-2).

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The authors declare that they have no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Yu, Y., Yuan, Y. & Gao, M. Construction of an environmental safe Bacillus thuringiensis engineered strain against Coleoptera . Appl Microbiol Biotechnol 100, 4027–4034 (2016). https://doi.org/10.1007/s00253-015-7250-5

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  • DOI: https://doi.org/10.1007/s00253-015-7250-5

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