期刊
JOURNAL OF MICROBIOLOGICAL METHODS
卷 99, 期 -, 页码 27-34出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2014.01.015
关键词
Soil; DNA extraction; Functional metagenomics; Broad-host-range cosmids; Metagenomic libraries; Gateway (R) recombination vector
资金
- Strategic Projects grant from the Natural Sciences and Engineering Research Council of Canada (NSERC)
We constructed IncP broad-host-range Gateway (R) entry cosmids pJC8 and pJC24, which replicate in diverse Proteobacteria. We demonstrate the functionality of these vectors by extracting, purifying, and size-selecting metagenomic DNA from agricultural corn and wheat soils, followed by cloning into pJC8. Metagenomic DNA libraries of 8 x 10(4) (corn soil) and 9 x 10(6) (wheat soil) clones were generated for functional screening. The DNA cloned in these libraries can be transferred from these recombinant cosmids to Gateway destination vectors for specialized screening purposes. Those library clones are available from the Canadian MetaMicroBiome Library project (http://www.cm2bl.org/). (C) 2014 Elsevier B.V. All rights reserved.
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