The bacmid system of BmMNPV with cysteine protease gene deletion (CPD-BmMNPV bacmid) was constructed using the lambda recombination system. The protease activities of Bombyx mori cells and silkworm larvae infected with this CPD-BmMNPV bacmid were reduced by 94% and 85%, respectively. By using this system, a GFP(uv)-beta1,3-N-acetylglucosaminyltransferase 2 (GFP(uv)-beta3GnT2) fusion protein was...

In the present study, we studied the feasibility of deleting essential genes in insect cells by using bacmid and purifying recombinant bacmid in Escherichia coli DH10B cells. To disrupt the orf4 (open reading frame 4) gene of BmNPV [Bm (Bombyx mori) nuclear polyhedrosis virus], a transfer vector was constructed and co-transfected with BmNPV bacmid into Bm cells. Three passages of viruses were c...

The Autographa californica multiple nucleocapsid nucleopolyhedrovirus (AcMNPV) alkaline nuclease (AN) associates with the baculovirus single-stranded DNA binding protein LEF-3 and possesses both a 5'-->3' exonuclease and an endonuclease activity. These activities are thought to be involved in DNA recombination and replication. To investigate the role of AN in AcMNPV replication, the lambda Red ...

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 to study the regulatory mechanism of DBP on viral proliferation, genome replication, and gene transcription, a BmNPV dbp gene knockout virus dbp-ko-Bacmid was generated by the me...

The most widely studied baculovirus, the Autographa californica M nucleopolyhedrovirus (AcMNPV), is a large, enveloped DNA virus that infects lepidopteran larvae (8). This virus is commonly used as a eukaryotic vector to express heterologous genes in cultured insect cells and insect larvae (1,3,15). Baculovirus expression systems offer several advantages over other expression systems, among whi...

Coronavirus disease (COVID-19) causes a serious threat to human health. Virus-like particles (VLPs) constitute promising platform in SARS-CoV-2 vaccine development. In this study, the E, M, and S genes were cloned into multiple cloning sites of new triple expression plasmid with one p10 promoter, two pPH promoters, three sites. The was transformed DH10 BacTMEscherichia coli competent cells obta...

Open reading frame 76 of Bombyx mori nucleopolyhedrovirus (BmNPV), designated as Bm76, is a gene whose function is completely unknown. With EGFP fused to the 3' terminal of Bm76 as the reporter gene and BmNPV bacmid as the expression vector, a recombinant bacmid was successfully constructed expressing Bm76-EGFP fusion protein under the control of polyhedrin promoter in Bombyx mori cells (Bm cel...

A Nicaraguan population of Spodoptera frugiperda multiple nucleopolyhedrovirus, SfMNPV-NIC, includes fast-killing genotypes with deletions in the egt region. Four bacmid based recombinants were constructed to determine the role of egt in this phenotype. SfdelF bacmid encompassed the deletion found in the NIC-F genotype. Sfdel3AP2 bacmid was constructed using the deletion reported in SfMNPV-3AP2...

Very late expression factor 1 (VLF-1) of Autographa californica multiple nucleopolyhedrovirus is a putative tyrosine recombinase and is required for both very late gene expression and budded virus production. In this report, we show that a vlf-1 knockout bacmid was able to synthesize viral DNA at levels similar to that detected for a gp64 knockout bacmid that served as a noninfectious control v...

BACKGROUND Crimean-Congo hemorrhagic fever virus (CCHFV) is a member of the nairovirus, a genus in the Bunyaviridae family, which causes a life threatening disease in human. Currently, there is no vaccine against CCHFV and detailed structural analysis of CCHFV proteins remains undefined. The CCHFV M RNA segment encodes two viral surface glycoproteins known as Gn and Gc. Viral glycoproteins can ...