Author: van der Schaar, H. M.; Melia, C. E.; van Bruggen, J. A. C.; Strating, J. R. P. M.; van Geenen, M. E. D.; Koster, A. J.; Bárcena, M.; van Kuppeveld, F. J. M.
Title: Illuminating the Sites of Enterovirus Replication in Living Cells by Using a Split-GFP-Tagged Viral Protein Document date: 2016_7_6
ID: 1aptufp6_16
Snippet: Subsequently, we tested whether the GFP(S11) tag in 3A affects the replication kinetics of the virus. Since cells that have been transfected with plasmid DNA are less susceptible to picornavirus infection (48), we generated single-cell clones of HeLa and BGM cells that stably express GFP(S1-10). Next, we compared the growth kinetics of CVB3-3A(S11aa2) to wild-type (wt) CVB3 in the presence and absence of GFP(S1-10). Samples were subjected to endp.....
Document: Subsequently, we tested whether the GFP(S11) tag in 3A affects the replication kinetics of the virus. Since cells that have been transfected with plasmid DNA are less susceptible to picornavirus infection (48), we generated single-cell clones of HeLa and BGM cells that stably express GFP(S1-10). Next, we compared the growth kinetics of CVB3-3A(S11aa2) to wild-type (wt) CVB3 in the presence and absence of GFP(S1-10). Samples were subjected to endpoint titration to determine the production of infectious virus or quantitative PCR to measure viral RNA levels. Replication of wt CVB3 in BGM cells and BGM(GFPS1-10) cells was nearly identical, showing that the presence of GFP(S1-10) did not alter CVB3 replication kinetics ( Fig. 2A and B) . The replication kinetics of tagged CVB3 in BGM(GFPS1-10) cells also resembled that of BGM cells,
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