Table 1.
Primer sequences for PCR amplification.
Fig 1.
Effect of dbp-knockout on viral replication.
(A) BmN cells transfected with dbp-ko-Bacmid, dbp-re-Bacmid or wtBacmid were harvested at 6–72 h p.t. and analyzed for gp41 by RT-qPCR for the viral genome copy number. (B) The BmN cells were transfected with the Bacmids, harvested at 0~72 h p.t., and analyzed for the virus titers. **p<0.01 vs wtBacmid and dbp-re-Bacmid. Values are expressed as means±SEM. Similar results were obtained in three independent experiments.
Fig 2.
RT-qPCR analysis of the transcriptional phases of dbp gene in cells infected with wtBacmid.
**P <0.01 the maximum transcription value vs the first detected value of dbp, lef-3, vp39, and p10.
Fig 3.
RT-qPCR analysis of the transcription levels of viral genes in cells transfected with Bacmids.
(A-C) early gene lef-3, ie-1 and dnapol. (D) late gene vp39. (E) very late gene p10. **p<0.01 vs wtBacmid and dbp-re-Bacmid.
Fig 4.
Western blot analysis of DBP and expression levels of viral proteins in BmN cells transfected with Bacmids.
(A-D) BmN cells transfected with dbp-ko-Bacmid, dbp-re-Bacmid, or wtBacmid were harvested at 48 h p.t. and analyzed by 6×his-tag monoclonal antibody or series of monoclonal antibodies DBP, LEF-3, VP39 and P10, respectively. Lanes M: protein markers; lane1: wtBacmid; lane 2: dbp-ko-Bacmid; lane 3: dbp-re-Bacmid.
Fig 5.
Electron microscopic analysis of cells transfected with the bacmids.
(A-C) BmN cells were analyzed at 24 h p.t. with wtBacmid: 5000×, 20000× and 50000×. (D-F) BmN cells were analyzed at 24 h p.t. with dbp-ko-Bacmid: 5000×, 20000× and 50000×. (G-I) BmN cells were analyzed at 24 h p.t. with dbp-re-Bacmid: 5000×, 20000× and 50000×. Nu, nucleus; Cy, cytoplasm; Nc, nucleocapsid. Bars indicate the scale.