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. 2004 Apr;48(4):1136–1144. doi: 10.1128/AAC.48.4.1136-1144.2004

FIG. 1.

FIG. 1.

Site of integration for ERG11 promoter fusion. The ERG11 promoter (black box) fused to the RLUC reporter gene (box with diagonal lines) was flanked by the backbone from plasmid pCRW3 (thin lines). PCRW3 also contains a functional ADE2 gene (shaded boxes). The plasmid containing the promoter was cut at the NsiI site for integration into the ADE2 genomic locus (white boxes) containing a deletion of part of the coding region (box with vertical stripes) that makes the CAI8 parental strain Ade−. The correct single integration is shown. Oligonucleotides were used in PCRs to screen for correct integrations of the construct. Oligonucleotides 2 (pSP73-2358) and 3 (ADE2-11487R) generate a DNA fragment that is a part of the plasmid pCRW3 backbone, which is a test for the presence of the construct in the transformants. Oligonucleotides 2 and 4 (ADE2-11545R) test for the correct integration of the ERG11 promoter construct into the ADE2 locus of CAI8. Oligonucleotide 1 (RLUC) was used as a probe for Southern blot analyses to make sure that a single copy of the ERG11 promoter construct was integrated into the ADE2 locus. The figure shows the linear order of the genes and the relative positions of the oligonucleotides but is not drawn to scale.