FIG. 6.

Eukaryotic expression analysis of the UL26 protein after infection of HFFs with HCMV strain AD169 or after transfection of COS-7 cells. (A) Western blot analysis with specific anti-UL26 antiserum and cell extracts from HFFs harvested at 72 h after infection with HCMV. Lanes: 1, extracts from HFFs 72 hpi with HCMV; 2, extracts from mock-infected HFFs; 3, prokaryotically expressed UL26 protein. (B) Western blot analysis with anti-UL26 antiserum and protein extracts harvested at various times after infection of human fibroblast cells with HCMV. Lanes: 1, mock-infected cells; 2, 6 hpi; 3, 24 hpi; 4, 30 hpi; 5, 48 hpi; 6, 72 hpi. (C) Western blot analysis with anti-UL26 antiserum and protein extracts from infected HFFs and transfected COS-7 cells. Lanes: 1, extracts from HFFs, 118 hpi; 2, extracts from COS-7 cells transfected with UL26 expression vector pHM266; 3, extracts from COS-7 cells transfected with expression vector pCB6; 4, prokaryotically expressed UL26. (D) Western blot analysis with anti-UL26 antiserum and extracts from transfected COS-7 cells. Lanes: 1, extracts from COS-7 cells that were transfected with the vector pCB6; 2, extracts from COS-7 cells that were transfected with UL26 expression plasmid pHM266; 3, extracts from COS-7 cells that were transfected with UL26 expression plasmid pHM619; 4, extracts from HFFs at 72 hpi with HCMV; 5, prokaryotically expressed pUL26. (E) Analysis of in vitro-translated proteins by SDS-PAGE and autoradiography. Lanes: 1, no template was used in the in vitro transcription-translation reaction; 2, the vector pcDNA3 was used in the in vitro transcription-translation reaction; 3, plasmid pHM1773 (containing both ATGs of UL26) was used in the in vitro transcription-translation reaction; 4, plasmid pHM1774 (containing only the predicted ATG of UL26) was used in the in vitro transcription-translation reaction. Sizes of the molecular mass markers are shown on the left of each panel; the sizes of pUL26 proteins are indicated on the right of panels A, B, D, and E.