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. 2002 Mar;76(5):2329–2339. doi: 10.1128/jvi.76.5.2329-2339.2002

FIG. 5.

FIG. 5.

Reverse transcription assays with vRNA-tRNA complexes isolated from wild-type and mutant virions. The amount of vRNA was quantitated by DNA primer extension with the poly(A) primer, which produced a 104-nt product (lanes 1 to 6). Extension of the PBS-bound tRNA primer resulted in a 257-nt tRNA-cDNA product on the wild-type (wt) template (lanes 7 to 12). The products are shorter on the d1 and d1/2 template due to the d1 deletion in the U5 region. A major RT pause product with a length of 186 nt is visible on the wild-type template. The PBS-tRNA occupancy was determined by extension of the downstream AUG primer (lanes 13 to 18). When reverse transcription is blocked by the annealed tRNA primer, a 175-nt cDNA is produced. Free RNA templates will produce a full-length cDNA product of 374 nt. An additional stop product is observed on the 2L and d1 templates (lanes 13 and 16; stop). This stop is partially resolved on the d1 rev1 template (lane 17).