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Improving conjugation efficacy of Sorangium cellulosum by the addition of dual selection antibiotics

  • Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The conjugation protocols in myxobacterium Sorangium cellulosum are often inapplicable due to the strain-specific sensitivity to the presence of Escherichia coli cells or the resistances to many antibiotics. Here we report that the conjugative transfer of the mobilizable plasmid pCVD442 from E. coli DH5α (λ pir) to Sorangium strains could be greatly increased by the presence of low doses of dual selection antibiotics in the mating medium. The improvement was efficient in either E. coli-tolerant or sensitive Sorangium strains. For those phleomycin and hygromycin tolerant Sorangium strains, chloramphenicol-resistance gene was developed as a new selectable marker by driving the resistance gene with the aphII promoter. Using the improved protocol, the epothilone biosynthetic pathway was inactivated by an insertion mutation in the biosynthetic genes of the producing Sorangium strains.

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Acknowledgments

The work was financially supported by grants 39870003, 30270023, 30370792, and 30400009 from the Chinese National Natural Science Foundation. Thanks to Dr. Edward C. Mignot of Shandong University for linguistic advice.

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Authors and Affiliations

  1. State Key Laboratory of Microbial Technology, College of Life Science, Shandong University, 250100, Jinan, People’s Republic of China

    Zhi-Jie Xia, Jing Wang, Wei Hu, Hong Liu, Xiu-Zhen Gao, Zhi-Hong Wu, Peng-Yi Zhang & Yue-Zhong Li

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  1. Zhi-Jie Xia
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  2. Jing Wang
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  3. Wei Hu
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  4. Hong Liu
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Correspondence to Yue-Zhong Li.

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Z.-J. Xia and J. Wang contributes equally to this work.

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Xia, ZJ., Wang, J., Hu, W. et al. Improving conjugation efficacy of Sorangium cellulosum by the addition of dual selection antibiotics. J Ind Microbiol Biotechnol 35, 1157–1163 (2008). https://doi.org/10.1007/s10295-008-0395-9

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  • DOI: https://doi.org/10.1007/s10295-008-0395-9

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