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Artificial induction of genetic competence in Bacillus amyloliquefaciens isolates

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Abstract

Objectives

To induce natural genetic competence in Bacillus amyloliquefaciens isolates through overexpression of the master regulator, ComK, from B. subtilis (ComK Bsu ).

Results

Plasmid pUBXC carrying the xylose-inducible comK expression cassette was constructed using plasmid pUB110 as a backbone. Plasmid pUBXC could be transferred from B. subtilis to B. amyloliquefaciens through plasmid pLS20-mediated biparental conjugation. After being induced by xylose, four B. amyloliquefaciens strains harbouring plasmid pUBXC developed genetic competence. Under optimal conditions, the transformation efficiencies of plasmid DNA ranged from 129 ± 20.6 to 1.7 ± 0.1 × 105 cfu (colony-forming units) per μg DNA, and the transformation efficiencies of PCR-assembled deletion constructs ranged from 3.2 ± 0.76 to 3.5 ± 0.42 × 104 cfu per μg DNA in the four tested strains.

Conclusion

Artificial induction of genetic competence through overexpressing ComK Bsu in B. amyloliquefaciens completed the tasks of replicative plasmid delivery and gene knockout via direct transformation of PCR-generated deletion cassettes.

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Acknowledgments

We appreciate Mitsuhiro Itaya from Mitsubishi Kagaku Institute of Life Science for providing plasmid pLS20cat. The authors would like to express their thanks to the financial support from 863 Plan (2014AA020543), 973 Plan (2015CB150505) and National Natural Science Foundation (31300099 and 31470225) of China.

Supporting information

Supplementary Table 1—Oligonucleotides used.

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Authors and Affiliations

  1. Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China

    Xue-Ting Chen, Jun-Bin Ji, Yong-Chuang Liu, Bin Ye, Chao-Yang Zhou & Xin Yan

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  1. Xue-Ting Chen
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  2. Jun-Bin Ji
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  3. Yong-Chuang Liu
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  4. Bin Ye
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  5. Chao-Yang Zhou
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  6. Xin Yan
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Corresponding author

Correspondence to Xin Yan.

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Chen, XT., Ji, JB., Liu, YC. et al. Artificial induction of genetic competence in Bacillus amyloliquefaciens isolates. Biotechnol Lett 38, 2109–2117 (2016). https://doi.org/10.1007/s10529-016-2194-0

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  • DOI: https://doi.org/10.1007/s10529-016-2194-0

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