Author: Stewart, Meredith E.; Roy, Polly
Title: Structure-based identification of functional residues in the nucleoside-2'-O-methylase domain of Bluetongue virus VP4 capping enzyme Document date: 2015_2_24
ID: vzel6r43_43
Snippet: To investigate the importance of 2 0 -O MT activity on virus replication we examined the D265 mutants in the context of virus replication. To this end the D265E and D265V mutations were introduced into the S4 and BTV reverse genetics (RG) system used [3, 19] to recover mutant viruses in mammalian BSR cells, WT VP4 plasmid was excluded from the first transfection and replaced with mutant T7 S4 ssRNA. In spite of several attempts, it was not possib.....
Document: To investigate the importance of 2 0 -O MT activity on virus replication we examined the D265 mutants in the context of virus replication. To this end the D265E and D265V mutations were introduced into the S4 and BTV reverse genetics (RG) system used [3, 19] to recover mutant viruses in mammalian BSR cells, WT VP4 plasmid was excluded from the first transfection and replaced with mutant T7 S4 ssRNA. In spite of several attempts, it was not possible to recover any virus, with either mutants of D265 in normal BSR cells, indicating that these substitution mutations were not tolerated and lethal for virus growth in mammalian cells (Fig. 6A) .
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