scholarly journals Efficient Expression and Processing of Ebola Virus Glycoprotein Induces Morphological Changes in BmN Cells but Cannot Rescue Deficiency of Bombyx Mori Nucleopolyhedrovirus GP64

Viruses ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 1067 ◽  
Author(s):  
Jinshan Huang ◽  
Na Liu ◽  
Fanbo Xu ◽  
Ellen Ayepa ◽  
Charles Amanze ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Signal Peptide ◽  
Ebola Virus ◽  
Virus Disease ◽  
Early Stage ◽  
Morphological Changes ◽  
Expression System ◽  
Bombyx Mori Nucleopolyhedrovirus ◽  
Native Signal Peptide ◽  
Bmn Cells

Ebola virus (EBOV) disease outbreaks have resulted in many fatalities, yet no licensed vaccines are available to prevent infection. Recombinant glycoprotein (GP) production may contribute to finding a cure for Ebola virus disease, which is the key candidate protein for vaccine preparation. To explore GP1,2 expression in BmN cells, EBOV-GP1,2 with its native signal peptide or the GP64 signal peptide was cloned and transferred into a normal or gp64 null Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid via transposition. The infectivity of the recombinant bacmids was investigated after transfection, expression and localization of EBOV-GP were investigated, and cell morphological changes were analyzed by TEM. The GP64 signal peptide, but not the GP1,2 native signal peptide, caused GP1,2 localization to the cell membrane, and the differentially localized GP1,2 proteins were cleaved into GP1 and GP2 fragments in BmN cells. GP1,2 expression resulted in dramatic morphological changes in BmN cells in the early stage of infection. However, GP1,2 expression did not rescue GP64 deficiency in BmNPV infection. This study provides a better understanding of GP expression and processing in BmN cells, which may lay a foundation for EBOV-GP expression using the BmNPV baculovirus expression system.

scholarly journals Construction of a novel secretion expression system guided by native signal peptide of PhoD inZymomonas mobilis

2014 ◽  
Vol 78 (4) ◽  
pp. 708-713 ◽  
Author(s):  
Bo Wu ◽  
Ming-Xiong He ◽  
Hong Feng ◽  
Zong-Xia Shui ◽  
Xiao-Yu Tang ◽  
...  
Keyword(s):  
Signal Peptide ◽  
Expression System ◽  
Secretion Expression ◽  
Native Signal Peptide

Bombyx mori nucleopolyhedrovirus utilizes a clathrin and dynamin dependent endocytosis entry pathway into BmN cells

2018 ◽  
Vol 253 ◽  
pp. 12-19 ◽  
Author(s):  
Min Feng ◽  
Jianjia Zhang ◽  
Weifan Xu ◽  
Haiping Wang ◽  
Xiangshuo Kong ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Bombyx Mori Nucleopolyhedrovirus ◽  
Entry Pathway ◽  
Bmn Cells

Proliferation of Bombyx mori nucleopolyhedrovirus strain H4 in BmN cells is enhanced by exchange of the F gene sequence with type strain T3

2021 ◽  
Vol 291 ◽  
pp. 198195
Author(s):  
Mami Sakai ◽  
Satoshi Kakutani ◽  
Shin-ichiro Asano ◽  
Masanao Sato ◽  
Hisanori Bando
Keyword(s):  
Bombyx Mori ◽  
Type Strain ◽  
Gene Sequence ◽  
Bombyx Mori Nucleopolyhedrovirus ◽  
F Gene ◽  
Bmn Cells

Expression of EGFP driven by BmNPV orf4 promoter, a novel immediate early promoter

Biologia ◽  
2009 ◽  
Vol 64 (2) ◽  
Author(s):  
Wu-Jie Su ◽  
Bing Li ◽  
Wei-De Shen ◽  
Yan Wu ◽  
Shan-Ying Zhu ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Fluorescent Protein ◽  
Early Stage ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Immediate Early ◽  
Early Promoter ◽  
Polyhedrin Promoter ◽  
Foreign Genes ◽  
Green Fluorescent

AbstractBombyx mori nucleopolyhedrovirus (BmNPV) orf4 has been shown to be expressed at very early stage of Bm-NPV infection cycle. In this study, using transient expression experiment, we demonstrated for the first time that orf4 promoter is an immediate early promoter, indicating that orf4 may play a role in the immediate-early stage of BmNPV infection. Moreover, with the recently developed Bac-to-Bac/BmNPV baculovirus expression system and a modified pFast-Bac1 whose polyhedrin promoter was replaced with orf4 promoter, a recombinant bacmid baculovirus expressing enhanced green fluorescent protein (EGFP) under the control of orf4 promoter in Bombyx mori (Bm) cells was successfully constructed. The result not only showed that the polyhedrin promoter can be replaced easily with other promoters to direct the expression of foreign genes by using this novel system but also laid the foundation for the rescue experiment of orf4 deletion mutant.

scholarly journals Evidence for Nucleic Acid Binding Ability and Nucleosome Association of Bombyx mori Nucleopolyhedrovirus BRO Proteins

2000 ◽  
Vol 74 (15) ◽  
pp. 6784-6789 ◽  
Author(s):  
Evgueni A. Zemskov ◽  
WonKyung Kang ◽  
Susumu Maeda
Keyword(s):  
Amino Acid ◽  
Dna Binding ◽  
Bombyx Mori ◽  
Binding Proteins ◽  
Dna Binding Proteins ◽  
Amino Acid Sequences ◽  
Micrococcal Nuclease ◽  
Nucleic Acid Binding ◽  
Bombyx Mori Nucleopolyhedrovirus ◽  
Bmn Cells

ABSTRACT The Bombyx mori nucleopolyhedrovirus (BmNPV) genome contains five related members of the bro gene family, all of which are actively expressed in infected BmN cells. Although their functions are unknown, their amino acid sequences contain a motif found in all known viral and prokaryotic single-stranded DNA binding proteins. To determine if they bind to nucleic acids, we fractionated the nuclei of BmNPV-infected BmN cells using a histone extraction protocol. We detected BRO-A, BRO-C, and BRO-D in the histone H1 fraction using anti-BRO antibodies. Micrococcal nuclease treatment released these BRO proteins from the chromatin fraction, suggesting their involvement in nucleosome structures. Chromatographic fractionation showed that BRO-A and/or BRO-C interacted with core histones. Expression of partial sequences of BRO-A proved that the N-terminal 80 amino acid residues were required for DNA binding activity. We also demonstrated that BmNPV BRO proteins underwent phosphorylation and ubiquitination followed by proteasome degradation, which may explain their distribution in the cytoplasm as well as the nucleus. We propose that BRO-A and BRO-C may function as DNA binding proteins that influence host DNA replication and/or transcription.

Purification of human papillomavirus-like particles expressed in silkworm using a Bombyx mori nucleopolyhedrovirus bacmid expression system

2018 ◽  
Vol 1096 ◽  
pp. 39-47 ◽  
Author(s):  
Robert Minkner ◽  
Rina Baba ◽  
Yae Kurosawa ◽  
Shinichiro Suzuki ◽  
Tatsuya Kato ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Bombyx Mori ◽  
Expression System ◽  
Bombyx Mori Nucleopolyhedrovirus

scholarly journals Highly Efficient Extracellular Production of Recombinant Streptomyces PMF Phospholipase D in Escherichia coli

Catalysts ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1057
Author(s):  
Jing Wang ◽  
Sheng Xu ◽  
Yang Pang ◽  
Xin Wang ◽  
Kequan Chen ◽  
...  
Keyword(s):  
Phospholipase D ◽  
Signal Peptide ◽  
Expression System ◽  
Signal Peptides ◽  
Extracellular Production ◽  
Secretory Expression ◽  
E Coli ◽  
Fusion Signal ◽  
Optimized Conditions ◽  
Native Signal Peptide

To achieve efficient bio-production of phospholipase D (PLD), PLDs from different organisms were expressed in E.coli. An efficient secretory expression system was thereby developed for PLD. First, PLDs from Streptomyces PMF and Streptomyces racemochromogenes were separately over-expressed in E.coli to compare their transphosphatidylation activity based on the synthesis of phosphatidylserine (PS), and PLDPMF was determined to have higher activity. To further improve PLDPMF synthesis, a secretory expression system suitable for PLDPMF was constructed and optimized with different signal peptides. The highest secretory efficiency was observed when the PLD * (PLDPMF with the native signal peptide Nat removed) was expressed fused with the fusion signal peptide PelB-Nat in E. coli. The fermentation conditions were also investigated to increase the production of recombinant PLD and 10.5 U/mL PLD was ultimately obtained under the optimized conditions. For the application of recombinant PLD to PS synthesis, the PLD properties were characterized and 30.2 g/L of PS was produced after 24 h of bioconversion when 50 g/L phosphatidylcholine (PC) was added.

scholarly journals Bioinformatic Analysis of the Human Recombinant Iduronate 2-Sulfate Sulfatase

2016 ◽  
Vol 10 (1) ◽  
pp. 124-132 ◽  
Author(s):  
Edwin D. Morales-Álvarez ◽  
Claudia M. Rivera-Hoyos ◽  
Patricia Landázuri ◽  
Raúl A. Poutou-Piñales ◽  
Aura M. Pedroza-Rodríguez
Keyword(s):  
Signal Peptide ◽  
Recombinant Dna ◽  
Expression System ◽  
3D Structure ◽  
Bioinformatic Analysis ◽  
Biochemical Properties ◽  
Enzyme Replacement ◽  
The Past ◽  
Human Enzyme ◽  
Native Signal Peptide

Mucopolysaccharidosis type II is a human recessive disease linked to the X chromosome caused by deficiency of lysosomal enzyme Iduronate 2-Sulfate Sulfatase (IDS), which leads to accumulation of glycosaminoglycans in tissues and organs. The human enzyme has been expressed inEscherichia coliandPichia pastorisin attempt to develop more successful expression systems that allow the production of recombinant IDS for Enzyme Replacement Therapy (ERT). However, the preservation of native signal peptide in the sequence has caused conflicts in processing and recognition in the past, which led to problems in expression and enzyme activity. With the main object being the improvement of the expression system, we eliminate the native signal peptide of human recombinant IDS. The resulting sequence showed two modified codons, thus, our study aimed to analyze computationally the nucleotide sequence of theIDSnhwithout signal peptide in order to determine the 3D structure and other biochemical properties to compare them with the native human IDS (IDSnh). Results showed that there are no significant differences between both molecules in spite of the two-codon modifications detected in the recombinant DNA sequence.

scholarly journals Two Cholesterol Recognition Amino Acid Consensus Motifs of GP64 with Uncleaved Signal Peptide Are Required for Bombyx mori Nucleopolyhedrovirus Infection

2021 ◽  
Vol 9 (3) ◽  
Author(s):  
Bifang Hao ◽  
Wenbin Nan ◽  
Ying Xu ◽  
Lin Liu ◽  
Na Liu ◽  
...  
Keyword(s):  
Amino Acid ◽  
Fusion Protein ◽  
Membrane Fusion ◽  
Bombyx Mori ◽  
Signal Peptide ◽  
Bombyx Mori Nucleopolyhedrovirus ◽  
Membrane Fusion Protein ◽  
Consensus Motifs

BmNPV is a severe pathogen that mainly infects silkworms. GP64 is the key membrane fusion protein that mediates BmNPV infection, and some studies have indicated that cholesterol and lipids are involved in BmNPV infection.

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