Fig. 2

Structural map of the PfKch1-GFP and GFP-PfKch2 expression plasmids. CYC-GAL10P, a hybrid promoter carrying the non-translated leader of the cytochrome-1 gene fused to the GAL10 upstream activating sequence; TEV, a Tobacco Etch Virus cleavage site; GFP-His₈, yeast enhanced GFP cDNA with eight subsequent histidine codons; 2µ, yeast 2 micron origin of replication; leu2-d, a poorly expressing allele of the β-isopropylmalate dehydrogenase gene; bla, a β-lactamase gene; pMB1, the pMB1 origin of replication; URA3, the yeast orotidine-5′-phosphate decarboxylase gene. Expression plasmid construction was done by insertion of either PfKch1/PfKch2 PCR fragments or PfKch1/PfKch2 and GFP PCR fragments into the linearized expression vector pEMBLyex4 by in vivo homologous recombination in S. cerevisiae. Regions used for homologous recombination are shown in pink and grey respectively