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. 2005 Nov 21;579(30):6699–6704. doi: 10.1016/j.febslet.2005.10.065

Figure 1.

figure image

Construction of the rAd‐VSPs and the evaluation of rAd‐mediated transduction and expression. (A) A schematic diagram showing the organization of the bicistronic expression cassette of the modified shuttle vector (pShuttle‐CMV‐GOI‐IRES‐eGFP) (upper panel) used for rAd construction and the cloned coding regions of SARS‐CoV VSPs, including S, S1, S2, E, M and N (lower panel). The amino acids were numbered according to the corresponding proteins of SARS‐CoV strain HK‐39. The amino acid sequence of SPpGH is shown in the key and the detailed sequence information of the IRES‐eGFP fragment is available at “http://www.addgene.org/pgvec1?f=c&identifier=1736&cmd=findpl”. (B) Assessment of the optimal MOI for maximal transduction efficiency. The percentage of eGFP expressing cells was accessed by flow cytometer with at least 1 × 105 cells were counted for each sample. Each data point of the three assays were determined in triplicate and represents the average of three independent experiments ± standard error mean (S.E.M). (C) Expression of SARS‐CoV VSPs in Vero E6 cells. The expressed proteins were detected by using anti‐V5 antibody and the sizes of the molecular marker were shown on the left of each blot.