FIG. 2.
(A) Schematic summary of the construction of plasmid pUSM1006. The 5′ promoter (P) and 3′ terminator (T) regions of the corresponding GRE3 gene were amplified using oligonucleotide pairs ARpL-ARpR and ARtL-ARtR, respectively. The selectable markers bla and URA3 are indicated on the plasmid by striped boxes, and the GRE3 promoter (GRE3P) and terminator (GRE3T) sequences are indicated by open boxes. Sites for restriction endonucleases are indicated as follows: B, BamHI; E, EcoRI; H, HindIII; X, XbaI. (B) The plasmid pUSM1006 was linearized by cutting plasmid pUSM1006 with XbaI and integrating it into the genome by homologous recombination. The GRE3 allele was subsequently looped out by homologous recombination between the GRE3 terminator sequences. ORF, open reading frame.