Abstract
A common limitation for the identification of novel activities from functional (meta) genomic screens is the low number of active clones detected relative to the number of clones screened. Here we demonstrate that constructing libraries with strains known to produce bioactives can greatly enhance the screening efficiency, by increasing the “hit-rate” and unmasking multiple activities from the same bacterial source.
Highlights
Functional metagenomics, which includes the cloning of total DNA obtained from an environment into the host bacterium and screening the recombinant clones for a desired activity, is currently aMar
To assess the efficiency of our functional screens we constructed a fosmid library from the DNA of six marine bacterial isolates known to have antibacterial properties [15], expressed the library in E. coli and screened for activity against both bacteria and the nematode Caenorhabditis elegans
The relatively high hit rate observed in our study indicates that the scarcity of DNA encoding for bioactivities might be a significant limitation for metagenomic screens
Summary
Functional metagenomics, which includes the cloning of total DNA obtained from an environment into the host bacterium and screening the recombinant clones for a desired activity, is currently aMar. Functional metagenomics, which includes the cloning of total DNA obtained from an environment into the host bacterium and screening the recombinant clones for a desired activity, is currently a. Some of the successes of these functional screens are illustrated by the discovery of antibiotics, such as terragine. Functional screens have assisted in the understanding of the genomic bases of biosynthetic pathways underlying the production of bioactive compounds in single organisms. Burke et al (2007) [5] screened an Escherichia coli fosmid library constructed with genomic DNA from the marine bacterium P. tunicata, which is known for its ability to produce various bioactive compounds [6]. Clones producing the antifungal compound tambjamine were identified and a biosynthetic pathway was proposed based on the expressed genes required for tambjamine production [5]
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