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. 2014 Sep 18;10(9):e1004532. doi: 10.1371/journal.pgen.1004532

Figure 1. Cre-recombinase based assay for transcription errors in vivo.

Figure 1

(A) Two constructs based on the promoters for the GAL1 and HO genes that allow low level expression of a mutant variant of the Cre recombinase (cre-Y324C) that has a cysteine substitution for the active site tyrosine. Suppression of the A to G mutation can result from transcriptional misincorporation of an A at that position yielding transient Cre activity. (B) Transient Cre activity can be detected as recombination events that restore function to a HIS3 based reporter. (i) The interval near the normal HIS3 gene. (ii) A loxP site inserted into an artificial intron in HIS3 is still a functional gene. SD = splice donor, SA = splice acceptor. (iii) Insertion of the kanMX gene flanked by loxP sites into the intron results in loss of HIS3 function. (iv) Inversion of the C-terminal portion of the his3 gene renders it defective. (v) Cre-mediated inversion of the construct in iv results in a functional HIS3 gene.