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

Figure 1.

Figure 1.

Replicating retroviral vectors containing IRES with various numbers of A's in the A bulge. (A) DNA sequence spanning the 3′ of MLV env and EMCV IRES in Toca 511 that contains 7 A's in the A bulge. Lower-case letters indicate the 3′ of the env sequence; underlined capital letters indicate MluI (ACGCGT) and PsiI (TTATAA) restriction enzyme sites flanking the IRES sequence; underlined lower-case letters indicate the 7 A's in the A bulge; italicized capital letters indicate the start codon of a transgene. (B) Diagram of the A bulge in the J-K bifurcation domain in EMCV IRES incorporated into RRV expressing yCD2 or GFP. The native ATG8 (AUG in RNA) and ATG9 are underlined; enlarged and underlined sequence represents the A bulge in the J-K bifurcation domain; lower-case letters indicate the 5′ sequences in the pyrimidine-rich region in the 3′ IRES. (C) yCD2 protein expression from transiently transfected 293T cells. GAPDH detection was included as a loading control. Positive control (+) is lysate from U87-MG cells infected with RRV-yCD2 vector. EMCV, encephalomyocarditis virus; IRES, internal ribosome entry site; MLV, murine leukemia virus; NC, negative control from naïve cell lysates.