Microfluidic automated plasmid library enrichment for biosynthetic gene cluster discovery

Author:

Xu Peng1ORCID,Modavi Cyrus1,Demaree Benjamin12,Twigg Frederick3,Liang Benjamin1,Sun Chen1,Zhang Wenjun34,Abate Adam R14

Affiliation:

1. Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA, USA

2. UC Berkeley-UCSF Graduate Program in Bioengineering, University of California, San Francisco, CA, USA

3. Department of Chemical and Biomolecular Engineering, University of California Berkeley, Berkeley, CA, USA

4. Chan Zuckerberg Biohub, San Francisco, CA, USA

Abstract

Abstract Microbial biosynthetic gene clusters are a valuable source of bioactive molecules. However, because they typically represent a small fraction of genomic material in most metagenomic samples, it remains challenging to deeply sequence them. We present an approach to isolate and sequence gene clusters in metagenomic samples using microfluidic automated plasmid library enrichment. Our approach provides deep coverage of the target gene cluster, facilitating reassembly. We demonstrate the approach by isolating and sequencing type I polyketide synthase gene clusters from an Antarctic soil metagenome. Our method promotes the discovery of functional-related genes and biosynthetic pathways.

Funder

National Institutes of Health

Publisher

Oxford University Press (OUP)

Subject

Genetics

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