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Science China Life Sciences

, Volume 62, Issue 8, pp 1087–1095 | Cite as

Strategy for efficient cloning of biosynthetic gene clusters from fungi

  • Ruixin Li
  • ZiXin Li
  • Ke Ma
  • Gang Wang
  • Wei Li
  • Hong-Wei Liu
  • Wen-Bing YinEmail author
  • Peng ZhangEmail author
  • Xing-Zhong LiuEmail author
Research Paper

Abstract

Filamentous fungi are excellent sources for the production of a group of bioactive small molecules which are often called secondary metabolites (SMs). The advanced genome sequencing technology combined with bioinformatics analysis reveals a large number of unexplored biosynthetic gene clusters (BGCs) in the fungal genomes. To unlock this fungal SM treasure, many approaches including heterologous expression are being developed and efficient cloning of the BGCs is a crucial step to do this. Here, we present an efficient strategy for the direct cloning of fungal BGCs. This strategy consisted of Splicing by Overlapping Extension (SOE)-PCR and yeast assembly in vivo. By testing 14 BGCs DNA fragments ranging from 7 kb to 52 kb, the average positive rate was over 80%. The maximal insertion size for fungal BGC assembly was 52 kb. Those constructs could be used conveniently for the heterologous expression leading to the discovery of novel natural products. Thus, our results provide an efficient and quick method for the low cost direct cloning of fungal BGCs.

Keywords

biosynthetic gene clusters Saccharomyces cerevisiae homologous recombination DNA assembly 

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Copyright information

© Science China Press and Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Life SciencesUniversity of Science and Technology of ChinaHefeiChina
  2. 2.State Key Laboratory of Mycology, Institute of MicrobiologyChinese Academy of SciencesBeijingChina

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