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To determine whether only the p36 isoform is translated by internal ribosome entry two truncated versions of the DNA encoding the p36 5' UTR were introduced into the dicistronic vectors. The vector pRB5'F that contained the 5' UTR up to the p46 ORF (Figure 5a) had no IRES activity in the assay after transient transfection in HeLa cells (Figure 5b). The vector pRB3'F that contains only the sequence between the p46 AUG and the p36 AUG had approximately 75% of the IRES activity of the fulllength p36 5' UTR (13 times the activity attributable to readthrough compared to 17 times for the fulllength).
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