Figure 4

Complementation studies of six glycolytic gene deletions with pUDC074. A) On synthetic medium complemented with all amino acids except uracil (SM ura DO) all isolated spores from a single tetrad of a heterozygous diploid of the indicated gene can grow, proving the presence of the plasmid in all spores. Those spores were replicated to SM ura DO medium complemented with G418. Only spores containing the chromosomal deletion of the represented gene can grow due to the selection for the KanMX marker. Spores growing on both media confirmed the presence of a functional copy of the gene on pUDC074. B) Complementation study of HXK2 with pUDC074 in a strain incapable of phosphorylating glucose. Spot plates are shown (10,000, 1000, 100, 10 cells/μl). Introduction of the plasmid restored the ability to grow on glucose as the sole carbon source.