An Efficient Markerless Deletion System Suitable for the Industrial Strains of Streptomyces

Dong Jianxin; Wei Jiaxiu; Li Han; Zhao Shiyao; Guan Wenjun

doi:10.4014/jmb.2106.06083

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자료유형: 학술저널

저자정보: Dong Jianxin (Institute of Pharmaceutical Biotechnology and The Children’s Hospital Zhejiang University School of) Wei Jiaxiu (Institute of Pharmaceutical Biotechnology and The Children’s Hospital Zhejiang University School of) Li Han (Institute of Pharmaceutical Biotechnology and The Children’s Hospital Zhejiang University School of) Zhao Shiyao (Institute of Pharmaceutical Biotechnology and The Children’s Hospital Zhejiang University School of) Guan Wenjun (Institute of Pharmaceutical Biotechnology and The Children’s Hospital Zhejiang University School of)

저널정보: 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제31권 제12호

발행연도: 2021.12

수록면: 1,722 - 1,731 (10page)

DOI: 10.4014/jmb.2106.06083

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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.

#Industrial Streptomyces #indigoidine synthetase #large DNA fragment deletion #biosynthetic gene cluster

참고문헌 (31)

참고문헌 신청

Bérdy J / 2005 / Bioactive Microbial Metabolites / J. Antibiot 58:1~26

Komatsu M / 2010 / Genome-minimized Streptomyces host for the heterologous expression of secondary metabolism / Proc. Natl. Acad. Sci. USA 107:2646~2651

Liu R / 2018 / Streptomyces species : Ideal chassis for natural product discovery and overproduction / Metab. Eng 50:74~84

Musiol-Kroll EM / 2019 / Challenges and advances in genetic manipulation of filamentous actinomycetes–the remarkable producers of specialized metabolites / Nat. Prod. Rep 36:1351~1369

Hopwood DA / 1999 / Genetic recombination and strain improvement (Vol. 18) / J. Ind. Microbiol. Biotechnol 22:323~335

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