Primary sequence domains required for the retention of rotavirus VP7 in the endoplasmic reticulum

COS7 cells (10) were grown and transfected as described previously (31) . Transfected cells in 100-mm dishes were labeled with L[35S]methionine at a concentration of 150-200 0`Ci/ml for 4 h at 37°C 46 h after transfection. Cell lysates and media were immunoprecipitated as described previously (31) except that either rabbit polyclonal anti-VP'/or rabbit anti-human salivary Q-amylase antiserum was used and incubation was for 1-2 h at 4°C. To inhibit asparagine-linked glycosylation, cells were incubated in medium containing tonicamycin (Sigma Chemical Co.) at a final concentration of 5 0`g/ml, for 4 h before, as well as during, the 4-h labeling period, as described previously (31) .