FIG 2.

Mutations at R128 and R129 in the GKYLQRRLQ motif impair the expression of nsp2TF in the vaccinia virus-T7 expression system. HEK-293T cells were infected with vaccinia virus expressing T7 polymerase at an MOI of 10 and then transfected with pLnsp1β-2 constructs at 1 h postinfection. The cell lysates were harvested at 18 h posttransfection. (A) nsp2TF was immunoprecipitated by a polyclonal antibody (PAb-TF) that specifically recognizes the C-terminal region of nsp2TF using equal amounts of the cell lysate. Immunoprecipitated proteins were detected by WB using PAb-TF (top) and MAb 140-68 recognizing the common N-terminal region of nsp2-related proteins (bottom). (B) Western blot detection of the expression of nsp2 (top) and nsp1β (bottom) using specific MAbs.