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. 2016 Jan 26;27(2):59–70. doi: 10.1089/hgtb.2015.131

Figure 4.

Figure 4.

Protein expression level of yCD2 in RRV-IRES-yCD2 variants decreases because of expansion of the oligo A length in the A bulge. Protein expression of yCD2 in RRV-IRES-yCD2 variants was evaluated at infection cycle 7 (A) and cycle 10 (C) to correlate with expansion of the oligo A length in A bulge A observed in sequencing results. Vector stability analyzed by PCR from maximally infected cells at infection cycle 7 (B) and cycle 10 (D) is shown to detect deletion of IRES-yCD2 cassette and noted as an additional factor in some variants contributing to the reduction of yCD2 protein expression. DNA molecular marker (1 kb plus marker; Invitrogen) is included in the first lane of each gel. Arrow indicates the expected 1.2 kb PCR product containing the IRES-yCD2 cassette. Asterisk indicates a deletion of the IRES-yCD2 cassette. +, positive control using RRV-IRES-6A plasmid DNA as a template in PCR; NTC, no template control.