Sequence-based screening for putative polyketide synthase gene-harboring clones from a soil metagenome library

Cited 0 time in scopus
Metadata Downloads
Title
Sequence-based screening for putative polyketide synthase gene-harboring clones from a soil metagenome library
Author(s)
Sang Chun Ji; Dockyu Kim; Jung-Hoon Yoon; Tae Kwang Oh; Choong Hwan Lee
Bibliographic Citation
Journal of Microbiology and Biotechnology, vol. 16, no. 1, pp. 153-157
Publication Year
2006
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.
Keyword
polyketide synthasesequence-based screeningsoil metagenome
ISSN
1017-7825
Publisher
South Korea
Type
Article
Appears in Collections:
Division of Biomedical Research > Metabolic Regulation Research Center > 1. Journal Articles
Files in This Item:
  • There are no files associated with this item.


Items in OpenAccess@KRIBB are protected by copyright, with all rights reserved, unless otherwise indicated.