doi: 10.1038/nmeth1073.
Epub 2007 Jul 22.
High-throughput cloning and expression in recalcitrant bacteria
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- PMID: 17643108
- DOI: 10.1038/nmeth1073
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High-throughput cloning and expression in recalcitrant bacteria
Nat Methods.
2007 Sep.
Free article
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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