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. Author manuscript; available in PMC: 2013 Apr 25.
Published in final edited form as: Virology. 2012 Feb 10;426(1):60–65. doi: 10.1016/j.virol.2012.01.019

Figure 4. Sequences surrounding the 537 AUG mediated its by-pass in spliced RNA.

Figure 4

(A): The map shows a schematic of the GPV genome between the RNA initiation site at nt 492 and the splice donor at nt 814. The 537 AUG and the 650 AUG are shown, as well as the regions of the GPV genome substituted with heterologous AAV5 capsid regions sequence (stippled) in the various constructs analyzed. As described in the text, all inserts were constructed to maintain the ORF1 reading frame throughout the insertion, and to maintain the HA tag at nt 540, which is also indicated. In addition, they contained a C-terminal ORF1 HA tag at nt 2241, which does not appear on this map. (B): Below is shown an immunoblot of protein extracts from 293T transfected with the constructs indicated and as diagrammed above and further described in the text. The doubly-tagged wild-type RepCap clone (N/CHA), as well as the cDNA-like clone singly N-terminally HA-tagged at nt 540 (NHA-cDNA), were included as controls. Rep1, Rep2, and Rep3, indicating usage of the 537 AUG in spliced RNA, are shown.