FIG 4.

The major 5′ end of the 4.9-kb Ty1i RNA maps to nt 1000. (A) Cap-independent 5′ RACE was performed with poly(A)⁺ RNA from wild-type BY4742 and an isogenic spt3Δ mutant (DG2247). The number of 5′ termini was plotted against the Ty1H3 sequence, and that and the distribution of the termini are on the x and y axes, respectively. The tallest peak represents the total number of 5′ ends captured at nt 1000 and is shown in parentheses. (B) 5′ RACE cDNA libraries from the wild-type and spt3Δ strains mentioned above and a repopulated S. paradoxus strain (DG2634) were amplified using a universal primer mix and a Ty1-specific primer, GSP1_3389. The amplification reaction mixtures were separated by agarose gel electrophoresis to demonstrate the presence of cDNA products corresponding to the 5′ ends of the full-length (5.7-kb) Ty1 and the truncated (4.9-kb) Ty1i RNAs.