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. Author manuscript; available in PMC: 2009 Oct 25.
Published in final edited form as: Virology. 2008 Aug 30;380(2):276–284. doi: 10.1016/j.virol.2008.07.013

Fig. 9. YFV replicon constructs and comparison of the effects of S56 mutations of YFV replicon growth.

Fig. 9

(A) Schematic representation of YFV replicon construct (Jones et.al., 2005) where the NS5 S56A and S56D mutations were done and used to measured the luciferase activity. * indicates first 22 amino acids of capsid protein for RNA cyclization sequences; R. Luc, Renilla luciferase; 2a, 17 amino acid residue auto proteolytic peptide from foot and mouth disease virus; black box, NS1 signal sequence; gray box, nonstructural protein sequences; arrow indicates in NS5 where S56A and S56D mutations were done. (B) YFV replicon RNA was transfected into HEK293T cells and incubated over 24 hours. At various time points, cells were lysed and luciferase activity corresponding to translation of viral RNA was measured. Wild type virus (filled diamonds), a GAA polymerase-dead mutant (circles) were used as controls for S56A (triangles) and S56D (squares). Values were normalized to readings obtained at 2 hours post transfection to control for transfection efficiency.