Figure 2
From: Properties and use of novel replication-competent vectors based on Semliki Forest virus
Protein expression in BHK-21 cells transfected with replication-competent vectors with duplicated SG promoters. BHK-21 cells were transfected with equal amount of in vitro transcripts prepared from pSFV4-T21/5-d1EGFP, pSFV4-T26/20-d1EGFP, pSFV4-T37/17-d1EGFP, pSFV4-T99/45-d1EGFP, pSFV4-d1EGFP-M96, pSFV4-d1EGFP-M37, pSFV4-d1EGFP-M26 and pSFV4-d1EGFP-M21; cells transfected with transcripts from pSP6-SFV4 were used as controls. Cells were collected at 12 h post-transfection, lysed in Laemmli buffer, and subjected to SDS-PAGE in 12% gel; each line corresponds to material from 50,000 cells. Proteins were transferred to nitrocellulose filter, probed by corresponding polyclonal antisera, and visualized by ECL. Sections of the blots probed by anti-SFV-nsP1 antiserum (top), anti-EGFP antiserum (middle) and anti-SFV-capsid antiserum (bottom) are shown. The bands corresponding to detected proteins are indicated with arrows on the left; the names of replication-competent vectors are shown above the blot. SFV4 indicates cells transfected with infectious transcripts from pSP6-SFV4.