FIG. 2.

Southern blot analysis of CaUPC2 strains. DNA from UPC2 deletion strains and the parental was digested with BglII, run on an agarose gel, blotted onto a nitrocellulose membrane, and then probed with the radiolabeled UPC2 5′ KO oligonucleotide (see Materials and Methods). The blot shows the replacement of a UPC2 allele with URA3 in strain S-7 and then the replacement of the remaining allele with ARG4 in strain D-6, confirming the disruption of both UPC2 alleles. Strains EC-7 and EC-2 are single-allele reconstructions replacing either the URA3 or ARG4 locus with a UPC2-HIS1 fragment. The additional lower band in the EC-7 lane is the result of the plasmid constructs.