scholarly journals A Comprehensive Roadmap Towards Generation of Influenza B Reporter Assay Using a Single DNA Polymerase Based Cloning of Reporter RNA Construct

2021 ◽  
Author(s):  
Nandita Kedia ◽  
Saptarshi Banerjee ◽  
Arindam Mondal
Keyword(s):  
Rna Synthesis ◽  
Rna Virus ◽  
Assay System ◽  
Viral Rna ◽  
Host Protein ◽  
Influenza B Virus ◽  
Influenza B ◽  
Reporter Assay ◽  
Virus Propagation ◽  
B Virus

Mini-genome reporter assay is a key tool for conducting RNA virus research. But, procedural complications and lack of adequate literature pose major challenge towards developing these assay systems. Here we present a novel yet generic and simple cloning strategy for construction of influenza B virus reporter RNA template and describe extensive standardization of the reporter RNP/polymerase activity assay for monitoring viral RNA synthesis in infection free setting. Using this assay system, we, for the first time showed the effect of viral protein NS1 and host protein PKC-Delta upon influenza B virus RNA synthesis. Additionally, the assay system showed promising results in evaluating the efficacy of antiviral drugs targeting viral RNA synthesis and virus propagation. Together, this work offers a detailed protocol for standardization of influenza virus mini-genome assay and an excellent tool for screening of host factors and antivirals in a fast, user friendly and high throughput manner.

MEK inhibition impairs influenza B virus propagation without emergence of resistant variants

FEBS Letters ◽  
2004 ◽  
Vol 561 (1-3) ◽  
pp. 37-43 ◽  
Author(s):  
Stephan Ludwig ◽  
Thorsten Wolff ◽  
Christina Ehrhardt ◽  
Walter Jürgen Wurzer ◽  
Jens Reinhardt ◽  
...  
Keyword(s):  
Influenza B Virus ◽  
Influenza B ◽  
Virus Propagation ◽  
Mek Inhibition ◽  
B Virus

scholarly journals The N-Terminal Extension of the Influenza B Virus Nucleoprotein Is Not Required for Nuclear Accumulation or the Expression and Replication of a Model RNA

1998 ◽  
Vol 72 (6) ◽  
pp. 5307-5312 ◽  
Author(s):  
Mark P. Stevens ◽  
Wendy S. Barclay
Keyword(s):  
Influenza A Virus ◽  
Influenza A ◽  
Viral Rna ◽  
Nuclear Accumulation ◽  
Influenza B Virus ◽  
Virus Protein ◽  
Influenza B ◽  
Coding Region ◽  
B Virus ◽  
N Terminus

ABSTRACT The nucleoprotein (NP) of influenza B virus is 50 amino acids longer at the N-terminus than influenza A virus NP and lacks homology to the A virus protein over the first 69 residues. We have deleted the N-terminal 51 and 69 residues of the influenza B/Ann Arbor/1/66 virus NP and show that nuclear accumulation of the protein is unaffected. This indicates that the nuclear localization signal is not located at the extreme N terminus, as in influenza A virus NP. To determine if the N-terminal mutants could support the expression and replication of a model influenza B virus RNA, the genes encoding the subunits of the viral RNA-dependent RNA polymerase (PA, PB1, and PB2) were cloned. Coexpression of NP and the P proteins in 293 cells was found to permit the expression and replication of a transfected model RNA based on segment 4 of B/Maryland/59, in which the hemagglutinin-coding region was replaced by a chloramphenicol acetyltransferase gene. The expression and replication of the synthetic RNA were not affected by the replacement of NP with NP mutants lacking the N-terminal 51 or 69 residues, indicating that the N-terminal extension is not required for transcription or replication of the viral RNA. In addition, we report that the influenza B virus NP cannot be functionally replaced by type A virus NP in this system.

Efficient influenza B virus propagation due to deficient interferon-induced antiviral activity in MDCK cells

Vaccine ◽  
2011 ◽  
Vol 29 (41) ◽  
pp. 7125-7129 ◽  
Author(s):  
Timo Frensing ◽  
Claudius Seitz ◽  
Bjoern Heynisch ◽  
Corinna Patzina ◽  
Georg Kochs ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Mdck Cells ◽  
Influenza B Virus ◽  
Influenza B ◽  
Virus Propagation ◽  
B Virus

scholarly journals ACETYLSALICYLIC ACID (ASA) IMPAIRS INFLUENZA B VIRUS PROPAGATION AND DELAY APOPTOSIS IN INOCULATED A549 AND MDCK II CELL LINES.

2016 ◽  
Vol 4 (5) ◽  
pp. 1247-1257
Author(s):  
Khalid Bassiouny ◽  
◽  
Ibrahim El-Sayed ◽  
Aziz Nokaly ◽  
AhmedS. Abdelghani ◽  
...  
Keyword(s):  
Acetylsalicylic Acid ◽  
Cell Lines ◽  
Influenza B Virus ◽  
Influenza B ◽  
Virus Propagation ◽  
B Virus

scholarly journals Recurring Influenza B Virus Infections in Seals

2013 ◽  
Vol 19 (3) ◽  
pp. 511-512 ◽  
Author(s):  
Rogier Bodewes ◽  
Danny Morick ◽  
Gerrie de Mutsert ◽  
Nynke Osinga ◽  
Theo Bestebroer ◽  
...  
Keyword(s):  
Influenza B Virus ◽  
Influenza B ◽  
Virus Infections ◽  
B Virus

scholarly journals Molecular Epidemiology and Phylogenetic Analyses of Influenza B Virus in Thailand during 2010 to 2014

PLoS ONE ◽  
2015 ◽  
Vol 10 (1) ◽  
pp. e0116302 ◽  
Author(s):  
Nipaporn Tewawong ◽  
Kamol Suwannakarn ◽  
Slinporn Prachayangprecha ◽  
Sumeth Korkong ◽  
Preeyaporn Vichiwattana ◽  
...  
Keyword(s):  
Molecular Epidemiology ◽  
Phylogenetic Analyses ◽  
Influenza B Virus ◽  
Influenza B ◽  
B Virus

scholarly journals Mutation E48K in PB1 Polymerase Subunit Improves Stability of a Candidate Live Attenuated Influenza B Virus Vaccine

Vaccines ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 800
Author(s):  
Jongsuk Mo ◽  
Stivalis Cardenas-Garcia ◽  
Jefferson J. S. Santos ◽  
Lucas M. Ferreri ◽  
C. Joaquín Cáceres ◽  
...  
Keyword(s):  
Vaccine Candidate ◽  
The Elderly ◽  
Potential Interaction ◽  
Respiratory Pathogen ◽  
Influenza B Virus ◽  
Influenza B ◽  
Epitope Tag ◽  
Live Attenuated Vaccine ◽  
B Virus ◽  
Homologous Challenge

Influenza B virus (IBV) is a major respiratory pathogen of humans, particularly in the elderly and children, and vaccines are the most effective way to control it. In previous work, incorporation of two mutations (E580G, S660A) along with the addition of an HA epitope tag in the PB1 segment of B/Brisbane/60/2008 (B/Bris) resulted in an attenuated strain that was safe and effective as a live attenuated vaccine. A third attempted mutation (K391E) in PB1 was not always stable. Interestingly, viruses that maintained the K391E mutation were associated with the mutation E48K. To explore the contribution of the E48K mutation to stability of the K391E mutation, a vaccine candidate was generated by inserting both mutations, along with attenuating mutations E580G and S660A, in PB1 of B/Bris (B/Bris PB1att 4M). Serial passages of the B/Bris PB1att 4M vaccine candidate in eggs and MDCK indicated high stability. In silico structural analysis revealed a potential interaction between amino acids at positions 48 and 391. In mice, B/Bris PB1att 4M was safe and provided complete protection against homologous challenge. These results confirm the compensatory effect of mutation E48K to stabilize the K391E mutation, resulting in a safer, yet still protective, IBV LAIV vaccine.

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