FIG. 7.

Detection of 5S integration using PCR. Top panel, yTM443 transformed with a low-copy-number plasmid expressing wild-type Ty3 (pMA1833) (upper panel) or Ty3 IN catalytic mutant (pMA1890) (lower panel) was induced for transposition in the presence of galactose for 6 h, and the DNA was harvested for PCR analysis. Ty3 elements integrated into the 5S rDNA were amplified using primers 411 (Ty3) and 720 (5S rDNA), and the PCR products were resolved by electrophoresis on a nondenaturing polyacrylamide gel and visualized by staining with ethidium bromide. Arrows indicate the positions of the amplified integration fragment. Negative control (N) contains only genomic DNA and primers. Positive control (P) contains genomic DNA, primers, and 0.05 ng of pDLC322, a plasmid containing a Ty3 upstream of a 5S rDNA gene (D. Chalker, unpublished data).