High-throughput cloning and expression in recalcitrant bacteriaGeertsma, E. R. & Poolman, B., Sep-2007, In : Nature Methods. 4, 9, p. 705-707 3 p.
Research output: Contribution to journal › Article › Academic › peer-review
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.
|Number of pages||3|
|Publication status||Published - Sep-2007|
- MEMBRANE-PROTEINS, STRUCTURAL GENOMICS, LACTOCOCCUS-LACTIS, RECOMBINANT-DNA, SYSTEMS