Public Library of Science
Browse
Figure_2.tif (530.75 kB)

rVSV-EnvG4-G6 characterization.

Download (0 kB)
figure
posted on 2014-09-12, 02:51 authored by Svetlana Rabinovich, Rebecca L. R. Powell, Ross W. B. Lindsay, Maoli Yuan, Alexei Carpov, Aaron Wilson, Mary Lopez, John W. Coleman, Denise Wagner, Palka Sharma, Marina Kemelman, Kevin J. Wright, John P. Seabrook, Heather Arendt, Jennifer Martinez, Joanne DeStefano, Maria J. Chiuchiolo, Christopher L. Parks

(a) After a 24 hr infection, total infected Vero cell lysates were collected and proteins were separated by SDS polyacrylamide gel electrophoresis (SDS-PAGE). Blot was probed with anti-VSV-N and anti-VSV-GIN CT, which does not recognize GNJ. IN: rVSV-EnvG4-G6IN; NJ: rVSV-EnvG4-G6NJ. (b-d) Sucrose-gradient purified rVSV-EnvG4-G6 particles were separated by SDS-PAGE. (b) Blots of 106 pfu of rVSV-EnvG4-G6IN and 2.5×106 pfu rVSV-EnvG4-G6NJ vectors were probed separately for EnvG using anti-gp120. (c) Blot of 106 pfu rVSV-EnvG4-G6IN was probed with anti-VSV-GIN CT. (d) Blot of 106 pfu rVSV-EnvG4-G6IN and rVSV-EnvG4-G6NJ was probed with anti-VSV-G. (e) Replication kinetics of rVSV-EnvG4-G6 and rVSV-G4 viruses in Vero cells. Vero cells were infected in 6-well plates at an MOI of 0.1. At various intervals post infection, supernatant was collected from duplicate wells and virus was titrated. Plaques were counted manually after cell fixation.

History