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Figure 1 | Virology Journal

Figure 1

From: Mutation of the elongin C binding domain of human respiratory syncytial virus non-structural protein 1 (NS1) results in degradation of NS1 and attenuation of the virus

Figure 1

Generation of rRSVs with a FLAG tag and mutations in NS1. (a) The full-length rRSV plasmid D46/6120 showing restriction enzyme sites used to introduce mutations (see Materials and Methods) and the position of a FLAG tag (grey bar) introduced to the N-terminus of NS1 (NS1F virus). Le = leader region. (b) Recovered NS1F virus was analysed by immunofluorescence in Vero cells by dual labelling with anti-FLAG monoclonal antibody (green) and anti-NS1/2 polyclonal antiserum (red). Nuclei were stained with DAPI and overlay images generated using Photoshop. (c) The 3 consensus residues of the NS1 elongin C binding domain (underlined) were replaced with alanine and the resulting cDNA was used to generate the NS1F-ELCmut virus. (d) Nucleotide substitutions (blue) that changed the 3 consensus residues to alanine in the recovered NS1F-ELCmut virus.

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Virology Journal

ISSN: 1743-422X