scholarly journals Generation of a recombinant rabies virus expressing green fluorescent protein for a virus neutralization antibody assay

2021 ◽  
Vol 22 ◽  
Author(s):  
Dong-Kun Yang ◽  
Ha-Hyun Kim ◽  
Yu-Ri Park ◽  
Jae Young Yoo ◽  
Yeseul Park ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Rabies Virus ◽  
Fluorescent Protein ◽  
Virus Neutralization ◽  
Antibody Assay ◽  
Green Fluorescent ◽  
Virus Neutralization Antibody

A new recombinant rabies virus expressing a green fluorescent protein: A novel and fast approach to quantify virus neutralizing antibodies

Biologicals ◽  
2019 ◽  
Vol 59 ◽  
pp. 56-61 ◽  
Author(s):  
Shuyun Qin ◽  
Dmitriy Volokhov ◽  
Elvira Rodionova ◽  
Christoph Wirblich ◽  
Matthias J. Schnell ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Rabies Virus ◽  
Neutralizing Antibodies ◽  
Fluorescent Protein ◽  
Protein A ◽  
Fast Approach ◽  
Green Fluorescent

A recombinant rabies virus expressing a phosphoprotein–eGFP fusion is rescued and applied to the rapid virus neutralization antibody assay

2015 ◽  
Vol 219 ◽  
pp. 75-83 ◽  
Author(s):  
Hai-Bo Tang ◽  
Zhuan-Ling Lu ◽  
Xian-Kai Wei ◽  
Yi-Zhi Zhong ◽  
Tao-Zhen Zhong ◽  
...  
Keyword(s):  
Rabies Virus ◽  
Virus Neutralization ◽  
Antibody Assay ◽  
Virus Neutralization Antibody

scholarly journals Tracking Fluorescence-Labeled Rabies Virus: Enhanced Green Fluorescent Protein-Tagged Phosphoprotein P Supports Virus Gene Expression and Formation of Infectious Particles

2004 ◽  
Vol 78 (22) ◽  
pp. 12333-12343 ◽  
Author(s):  
Stefan Finke ◽  
Krzysztof Brzózka ◽  
Karl-Klaus Conzelmann
Keyword(s):  
Gene Expression ◽  
Green Fluorescent Protein ◽  
Rabies Virus ◽  
Enhanced Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Live Cell ◽  
Fluorescence Labeling ◽  
Extracellular Virus ◽  
P Gene ◽  
Green Fluorescent

ABSTRACT Rhabdoviruses such as rabies virus (RV) encode only five multifunctional proteins accomplishing viral gene expression and virus formation. The viral phosphoprotein, P, is a structural component of the viral ribonucleoprotein (RNP) complex and an essential cofactor for the viral RNA-dependent RNA polymerase. We show here that RV P fused to enhanced green fluorescent protein (eGFP) can substitute for P throughout the viral life cycle, allowing fluorescence labeling and tracking of RV RNPs under live cell conditions. To first assess the functions of P fusion constructs, a recombinant RV lacking the P gene, SAD ΔP, was complemented in cell lines constitutively expressing eGFP-P or P-eGFP fusion proteins. P-eGFP supported the rapid accumulation of viral mRNAs but led to low infectious-virus titers, suggesting impairment of virus formation. In contrast, complementation with eGFP-P resulted in slower accumulation of mRNAs but similar infectious titers, suggesting interference with polymerase activity rather than with virus formation. Fluorescence microscopy allowed the detection of eGFP-P-labeled extracellular virus particles and tracking of cell binding and temperature-dependent internalization into intracellular vesicles. Recombinant RVs expressing eGFP-P or an eGFP-P mutant lacking the binding site for dynein light chain 1 (DLC1) instead of P were used to track interaction with cellular proteins. In cells expressing a DsRed-labeled DLC1, colocalization of DLC1 with eGFP-P but not with the mutant P was observed. Fluorescent labeling of RV RNPs will allow further dissection of virus entry, replication, and egress under live-cell conditions as well as cell interactions.

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