A novel method for rapid and sensitive metagenomic activity screening

Authors: Shang, Meiling; Chan, Victor; Wong, Dominic; HANS, LIAO - Cargill, Incorporated

Submitted to: MethodsX
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/10/2018
Publication Date: 6/20/2018
Citation: Shang, M., Chan, V.J., Wong, D., Hans, L. 2018. A novel method for rapid and sensitive metagenomic activity screening. MethodsX. 5:669-675. https://doi.org/10.1016/j.mex.2018.06.011.
DOI: https://doi.org/10.1016/j.mex.2018.06.011

Interpretive Summary: It has been estimated that less than 1% of the microbes in the natural environment can be cultured in laboratory conditions. In the last decade, direct cloning of the collective genomes of environmental microflora has been used as a powerful tool to explore the diversity and complexity of genes and enzymes in the unculturable microbial world. This approach requires development of rapid and sensitive high-throughput function-based screening of gene libraries. This report describes a rapid and sensitive screening method that could be universally adopted for discovery of novel genes from metagenomes with high efficacy.

Technical Abstract: Direct cloning of metagenomes has proven to be a powerful tool in the exploration of the diverse sequence space of a microbial community leading to recent advances in gene discovery and biocatalyst development. The key to such approach is the development of rapid, sensitive and reliable screening of libraries. A novel method is described, which includes: the formulation and application of substrate gel assay plates for rapid and sensitive screening of library of clones in split pools, and a progressive enrichment process and isolation of individual positive clones. The method has been applied in the discovery of numerous novel genes from various metagenomes with high efficacy.