Virus culture
MHV-68 expressing GFP (provided by R. Sun, University of California, Los
Angeles, CA) was passaged in NIH 3T3 cells with DMEM plus 10% FCS.
After lysis, supernatant was harvested, filtered (0.45-mm pore), and titered
by 2-fold serial dilutions. To determine virus load in infected mice, frozen
homogenized tissues were minced and subjected to 10-fold serial dilutions,
and endpoint titers were determined in NIH 3T3 cells by GFP (41, 42). A
single virion or DNA copy was sufficient to show a positive result by
plaque assay or PCR.
Virus cultureMHV-68 expressing GFP (provided by R. Sun, University of California, LosAngeles, CA) was passaged in NIH 3T3 cells with DMEM plus 10% FCS.After lysis, supernatant was harvested, filtered (0.45-mm pore), and titeredby 2-fold serial dilutions. To determine virus load in infected mice, frozenhomogenized tissues were minced and subjected to 10-fold serial dilutions,and endpoint titers were determined in NIH 3T3 cells by GFP (41, 42). Asingle virion or DNA copy was sufficient to show a positive result byplaque assay or PCR.
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