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Sequence-Based Screening for Putative Polyketide Synthase Gene-Harboring Clones from a Soil Metagenome Library  

JI SANG CHUN (Korea Research Institute of Bioscience and Biotechnology)
KIM DOCKYU (Korea Research Institute of Bioscience and Biotechnology)
YOON JUNG-HOON (Korea Research Institute of Bioscience and Biotechnology)
OH TAE-KWANG (Korea Research Institute of Bioscience and Biotechnology)
LEE CHOONG-HWAN (Korea Research Institute of Bioscience and Biotechnology)
Publication Information
Journal of Microbiology and Biotechnology / v.16, no.1, 2006 , pp. 153-157 More about this Journal
Abstract
A soil metagenomic library was constructed using an E. coli-fosmid cloning system with environmental DNAs extracted from Kwangreung forest topsoil. We targeted the genes involved in the biosynthesis of bacterial polyketides. Initially, a total of 36 clone pools (10,800 clones) were explored by the PCR-based method using the metagenomic DNAs from each pool and a degenerate primer set, which has been designed based on the highly conserved regions among ketoacyl synthase (KS) domains in actinomycete type I polyketide synthases (PKS Is). Six clone pools were tentatively selected as positive and further examined through a hybridization-based method for selecting a fosmid clone containing PKS I genes. Colony hybridization was performed against fosmid clones from the 6 positive pools, and finally 4 clones were picked out and confirmed to contain the conserved DNA fragment of KS domains. In this study, we present a simple and feasible sorting method for a desired clone from metagenomic libraries.
Keywords
Soil metagenome; sequence-based screening; polyketide synthase;
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Times Cited By KSCI : 3  (Citation Analysis)
Times Cited By Web Of Science : 5  (Related Records In Web of Science)
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