- Phenotype data
- Requesting materials
- Related resources
PlasmoGEM uses a combination of recombineering and Gateway® technology to convert genomic libraries into gene targeting vectors with long homology arms, which integrate with high efficiency (Pfander et al., 2011).
All PlasmoGEM vectors are equipped with sequence-readable barcodes, and we use barcode-sequencing (BarSeq) for high-throughput parallel growth phenotyping of mutants (method in Gomes et al., 2015).
The first step in the vector production pipeline uses recombineering (recombinase-mediated engineering; Zhang et al. 1998, Wang et al., 2006) and produces an intermediate vector in which a Zeo/PheS bacterial resistance marker is inserted into the coding sequence (gene knock-out) or 3'UTR (c-terminal tagging) of the target locus. The process is completed by a Gateway®-mediated exchange of the Zeo/PheS cassette for an hdhfr-yfcu parasite selectable marker.
Two types of gene targeting vectors are currently available from PlasmoGEM:
Final and intermediate PlasmoGEM gene targeting vectors, as well as toolkit vectors can be requested free of charge: