FIGURE 7.

3′-RACE of M. genitalium tRNA₁ ^Gly products. RNA was ligated to the DNA linker as described in Materials and Methods and in the legend of Figure 5. RT-PCR was carried out using primer pairs P1 (RP + tRNA-LP1), P2 (RP + tRNA-LP2), P3 (RP + tRNA-LP3), or a single primer RP. The expected PCR products from the mature 3′-end of tRNA₁ ^Gly using P1, P2, or P3 are labeled as M1, M2, or M3, respectively. A 25-bp DNA Step Ladder (25bp ladder; Promega) was used as the size marker in each gel. (A) RT-PCR products from total RNA isolated from M. genitalium culture (M.g. RNA). (B) PCR products using DNA templates gel-isolated from A: (Lane 2) DNA1 products above M1, up to ∼100 bp longer than M1; (lane 3) DNA2 products above M2, up to ∼100 bp longer than M2. Major PCR products longer than the products from mature tRNA are labeled Pre1, Pre2, and Pre3 on the right. (C) RT-PCR products from the 74-nt RNA species generated from treatment of pre-tRNA₁ ^Gly by MgR. Total RNA from M. genitalium (M.g. RNA) was used to show the sizes of PCR products from mature tRNA₁ ^Gly.