Figure 2.

Construction of GLY39, a new kex2 disrupted strain. (A) In M213 the KEX2 locus was only interrupted with the HIS3 marker, whereas in GLY39 the KEX2 locus was actually disrupted with the LEU2 marker. The thick arrow represents transcription direction of the LEU2 marker, and thin arrows indicate the hybridization site of oligonucleotides used for PCR. (B) Control of M213 and GLY39 growth on rich medium (YPD) and medium lacking histidine (−HIS) or leucine (−LEU). (C) Confirmation of GLY39 genotype. PCR was performed with genomic DNA from GLY39 (lanes 1 and 3) and the parental M213 strain (lanes 2 and 4). Negative controls without oligonucleotide are shown (lanes 3 and 4).