A DNA Binding Protein Is Required for Viral Replication and Transcription in Bombyx mori Nucleopolyhedrovirus

• DOI: 10.1371/journal.pone.0159149
• Publication Date: 2016-07-14T20:11:42Z
• Authors: Cui Zhao; Chen Zhang; Bin Chen; Yanghui Shi; Yanping Quan; Zuoming Nie; Yaozhou Zhang; Wei Yu
• Keywords: Transcription

A DNA-binding protein (DBP) [GenBank accession number: M63416] of Bombyx mori nuclear polyhedrosis virus (BmNPV) has been reported to be a regulatory factor in BmNPV, but its detailed functions remain unknown. In order study the mechanism DBP on viral proliferation, genome replication, and gene transcription, BmNPV dbp knockout dbp-ko-Bacmid was generated by means Red recombination system. addition, dbp-repaired dbp-re-Bacmid constructed Bac Then, Bacmids were transfected into BmN cells. The results this titer experiment revealed that TCID50 0; however, similar wtBacmid (p>0.05), indicating dbp-deficient would lead failure assembly particles. next step, Real-Time PCR used analyze transcriptional phases cells, which had infected with BmNPV. latter transcript first detected at 3 h post-infection. Furthermore, replication level early, late, very late genes kinds Bacmids, analyzed PCR. demonstrating lower than (p<0.01). early lef-3, ie-1, dnapol, vp39 p10 statistically significantly presented are based Western blot analysis, indicated lack low expressions lef3, vp39, p10. conclusion, not only essential for also significant impact transcription expression during all periods baculovirus life cycle.