Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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An Efficient Markerless Deletion System Suitable for the Industrial Strains of Streptomyces

  • Dong, Jianxin (Institute of Pharmaceutical Biotechnology and The Children's Hospital, Zhejiang University School of Medicine) ;
  • Wei, Jiaxiu (Institute of Pharmaceutical Biotechnology and The Children's Hospital, Zhejiang University School of Medicine) ;
  • Li, Han (Institute of Pharmaceutical Biotechnology and The Children's Hospital, Zhejiang University School of Medicine) ;
  • Zhao, Shiyao (Institute of Pharmaceutical Biotechnology and The Children's Hospital, Zhejiang University School of Medicine) ;
  • Guan, Wenjun (Institute of Pharmaceutical Biotechnology and The Children's Hospital, Zhejiang University School of Medicine)
  • Received : 2021.06.30
  • Accepted : 2021.09.02
  • Published : 2021.12.28
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Abstract

The genus Streptomyces is intensively studied due to its excellent ability to produce secondary metabolites with diverse bioactivities. In particular, adequate precursors of secondary metabolites as well as sophisticated post modification systems make some high-yield industrial strains of Streptomyces the promising chassis for the heterologous production of natural products. However, lack of efficient genetic tools for the manipulation of industrial strains, especially the episomal vector independent tools suitable for large DNA fragment deletion, makes it difficult to remold the metabolic pathways and streamline the genomes in these strains. In this respect, we developed an efficient deletion system independent of the episomal vector for large DNA fragment deletion. Based on this system, four large segments of DNA, ranging in length from 10 kb to 200 kb, were knocked out successfully from three industrial Streptomyces strains without any marker left. Notably, compared to the classical deletion system used in Streptomyces, this deletion system takes about 25% less time in our cases. This work provides a very effective tool for further genetic engineering of the industrial Streptomyces.

Keywords

Acknowledgement

This work was financially supported by National Key R&D Program of China (2019YFA0905400).

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