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High-throughput cloning and expression in recalcitrant bacteria

Geertsma, E. R. & Poolman, B., Sep-2007, In : Nature Methods. 4, 9, p. 705-707 3 p.

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DOI

We developed a generic method for high-throughput cloning in bacteria that are less amenable to conventional DNA manipulations. The method involves ligation-independent cloning in an intermediary Escherichia coli vector, which is rapidly converted via vector-backbone exchange (VBEx) into an organism-specific plasmid ready for high-efficiency transformation. We demonstrated VBEx proof of principle for Lactococcus lactis, but the method can be adapted to all organisms for which plasmids are available.

Original languageEnglish
Pages (from-to)705-707
Number of pages3
JournalNature Methods
Volume4
Issue number9
Publication statusPublished - Sep-2007

    Keywords

  • MEMBRANE-PROTEINS, STRUCTURAL GENOMICS, LACTOCOCCUS-LACTIS, RECOMBINANT-DNA, SYSTEMS

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