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  • Title: High-throughput cloning and expression in recalcitrant bacteria.
    Author: Geertsma ER, Poolman B.
    Journal: Nat Methods; 2007 Sep; 4(9):705-7. PubMed ID: 17643108.
    Abstract:
    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.
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