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Nat Methods. 2007 Sep;4(9):705-7. Epub 2007 Jul 22.

High-throughput cloning and expression in recalcitrant bacteria.

Author information

1
Department of Biochemistry, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG, Groningen, The Netherlands.

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.

PMID:
17643108
DOI:
10.1038/nmeth1073
[Indexed for MEDLINE]

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