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Is Transcription Interrupted by Transcriptional Terminators in Environmental Bacteria?

 To explore whether transcription of cloned metagenomic DNA is interrupted by transcriptional terminators in environmental bacteria, we employed Substrate Induced Gene Expression (SIGEX) technology24. This method involves mixing cultures of bacterial strains that stably carry the same fosmid and...

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 We have constructed a genetically compatible system that allows the transfer of metagenomic DNA fragments from rifampicin resistant (RifR) and nalidixic acid resistant (NalR) derivatives of E. coli to non-adapted host strains with very high efficiency. This system takes advantage of the specialized transcription systems of these host strains and their innate resistance to b-lactam antibiotics. This strategy was used to select for fosmids containing a gene encoding the components of an efflux...

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 The ability to efficiently clone metagenomic DNA and to identify active clones is a critical step towards the effective utilization of the extraordinary biodiversity of uncultivated bacteria. However, a significant limitation of current functional metagenomic screening systems is the inability to detect complex regulatory systems by identifying their transcriptional activators or anti-termination proteins. The MPO555 system overcomes this limitation by integrating a promoterless GFP gene downstream...

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