Journal: The Journal of General Virology

Article Title: An infectious recombinant foot-and-mouth disease virus expressing a fluorescent marker protein

doi: 10.1099/vir.0.052308-0

Figure Lengend Snippet: Determination of the packaging limitations for the targeted insertion site of the FMDV genome. (a) Schematic representation of the FMDV genome, showing the positions of the forward (F) and reverse (R) primers used to investigate the retention of different portions of the GFP gene. (b) Top: PCR controls performed using the T2- to T7-FMDV infectious copy plasmids as templates (see ). The PCR products show the expected amplification products derived from each of the FMDV genomes in the absence of deletions. Bottom: RT-PCR analysis of FMDV genomes (T2- to T7-FMDV) containing different portions of the GFP gene. FMDV genomes were prepared from goat epithelium cells infected with P0 virus stocks. T6- and T7-FMDV clearly show deletion of their respective insertions.

Article Snippet: A goat epithelium cell line expressing the principal FMDV receptor (integrin αvβ6) was subsequently used to passage the tagged viruses (P1) ( Brehm et al. , 2009 ).

Techniques: Amplification, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Infection, Virus

Journal: The Journal of General Virology

Article Title: An infectious recombinant foot-and-mouth disease virus expressing a fluorescent marker protein

doi: 10.1099/vir.0.052308-0

Figure Lengend Snippet: iLOV-FMDV is viable and stable. (a) Western blot analysis of whole-cell lysates prepared from goat epithelium cells infected with iLOV-FMDV. Samples were probed for the presence of iLOV, the non-structural FMDV 3A protein (and 3A-B precursors, black bar) and γ-tubulin (loading control). Asterisks indicate non-specific bands. (b) Fluorescence microscopy of goat epithelium cells infected with iLOV-FMDV. Nuclei are stained blue (DAPI). (c) RT-PCR analysis of iLOV-FMDV genomes prepared from P2, P3 and P4 virus stocks. Genome derived from the parental wild-type virus was also analysed as a control. (d) Growth analysis comparison of parental wild-type FMDV and iLOV-FMDV. Goat epithelium cells were infected with the respective virus (0.1 m.o.i.) and samples analysed at 0, 2, 4, 6 and 8 h post-infection. Similar results were obtained from three individual experiments. (e) Plaque morphology of goat epithelium cells infected with parental wild-type FMDV or iLOV-FMDV.

Article Snippet: A goat epithelium cell line expressing the principal FMDV receptor (integrin αvβ6) was subsequently used to passage the tagged viruses (P1) ( Brehm et al. , 2009 ).

Techniques: Western Blot, Infection, Control, Fluorescence, Microscopy, Staining, Reverse Transcription Polymerase Chain Reaction, Virus, Derivative Assay, Comparison

Journal: The Journal of General Virology

Article Title: An infectious recombinant foot-and-mouth disease virus expressing a fluorescent marker protein

doi: 10.1099/vir.0.052308-0

Figure Lengend Snippet: Detection of iLOV-FMDV-infected cells by flow cytometry. Goat epithelium cells were either mock infected or infected with iLOV-FMDV or parental FMDV at an m.o.i. of 2. After 4 h, cells were analysed by flow cytometry. (a–h) Density plots for iLOV-FMDV-infected or parental-FMDV-infected cells. (a, b) Infected and unlabelled cells analysed for iLOV expression. (c, d) Infected cells analysed for iLOV and FMDV non-structural protein 3A expression as detected with mAb 2C2. (e, f) Infected cells analysed for iLOV and FMDV capsid expression as detected with mAb IB11. (g, h) Infected cells analysed for iLOV expression and labelled with isotype control mAbs TRT1 and TRT3. (i–l) Histograms displayed for unlabelled cells infected with parental FMDV (i), mock infected (j), infected with iLOV-FMDV (k) and overlay of parental-FMDV-infected (red) and ILOV-FMDV-infected (black) cells (l).

Article Snippet: A goat epithelium cell line expressing the principal FMDV receptor (integrin αvβ6) was subsequently used to passage the tagged viruses (P1) ( Brehm et al. , 2009 ).

Techniques: Infection, Flow Cytometry, Expressing, Control

Journal: The Journal of General Virology

Article Title: An infectious recombinant foot-and-mouth disease virus expressing a fluorescent marker protein

doi: 10.1099/vir.0.052308-0

Figure Lengend Snippet: Detection of iLOV-FMDV-infected cells by live-cell imaging. (a–c) Still images from live-cell imaging experiments during which goat epithelium cells were infected at an m.o.i. of 2 with either parental-FMDV (a) or iLOV-FMDV (b) or mock infected (c). Cells imaged by differential interference contrast optics are shown. iLOV-expressing cells (green) are clearly visible in (b). Bar, 20 µm.

Article Snippet: A goat epithelium cell line expressing the principal FMDV receptor (integrin αvβ6) was subsequently used to passage the tagged viruses (P1) ( Brehm et al. , 2009 ).

Techniques: Infection, Live Cell Imaging, Expressing