Arrayed Plasmodium genomic libraries were constructed using the pJAZZ vector backbone. pJazz is a phage N15-derived vector that was once available from Lucigen (Godiska et al., 2010). pJAZZ is a low copy, linear vector especially suitable for maintaining large inserts of AT-rich DNA. The 14 kb long backbone of the vector contains a number of genetic features necessary for its propagation in the TSA strain of Escherichia coli. These features include hairpin telomers, a telomerase gene, replication factor and origin, and a selectable kanamycin resistance marker.
The gDNA inserts are end-sequenced and mapped to their genome of origin. The arrayed library clones are the starting point for recombineering. To search the gDNA libraries, tick the "library clones" box under "Filter for available PlasmoGEM resources" on the search form for genes. The result will show all genes that have overlapping library clones. The "lib clone" buttons in the gene result list show the library clones that are available.
We are currently making available library clones for Plasmodium berghei and Plasmodium chabaudi. More than 97% of the annotated gene models are covered (>50% of CDS) in P. berghei and >80% in P. chabaudi. The PbG libraries encompass 9,946 genomic clones, with insert sizes ranging from 4-20 kb and an average insert length of 8.7 kb.