Determination of cold-adapted influenza virus (Orthomyxoviridae: Alphainfluenzavirus ) polymerase activity by the minigenome method with a fluorescent protein.

Autor:	Ivanov PA; N.F. Gamaleya National Research Centre for Epidemiology and Microbiology, the Russian Ministry of Health., Lyashko AV; N.F. Gamaleya National Research Centre for Epidemiology and Microbiology, the Russian Ministry of Health., Kost VY; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences., Lomakina NF; N.F. Gamaleya National Research Centre for Epidemiology and Microbiology, the Russian Ministry of Health., Rtishchev AA; Mechnikov Research Institute for Vaccines and Sera., Bunkova NI; N.F. Gamaleya National Research Centre for Epidemiology and Microbiology, the Russian Ministry of Health., Timofeeva TA; N.F. Gamaleya National Research Centre for Epidemiology and Microbiology, the Russian Ministry of Health., Balanova MA; N.F. Gamaleya National Research Centre for Epidemiology and Microbiology, the Russian Ministry of Health., Ionov SA; N.F. Gamaleya National Research Centre for Epidemiology and Microbiology, the Russian Ministry of Health.; Mendeleev University of Chemical Technology., Gorikov DV; N.F. Gamaleya National Research Centre for Epidemiology and Microbiology, the Russian Ministry of Health.; Mendeleev University of Chemical Technology., Markushin SG; Mechnikov Research Institute for Vaccines and Sera.
Jazyk:	angličtina
Zdroj:	Voprosy virusologii [Vopr Virusol] 2023 Dec 26; Vol. 68 (6), pp. 526-535. Date of Electronic Publication: 2023 Dec 26.
DOI:	10.36233/0507-4088-203
Abstrakt:	Introduction: Polymerase proteins PB1 and PB2 determine the cold-adapted phenotype of the influenza virus A/Krasnodar/101/35/59 (H2N2), as was shown earlier. Objective: The development of the reporter construct to determine the activity of viral polymerase at 33 and 37 °C using the minigenome method. Materials and Methods: Co-transfection of Cos-1 cells with pHW2000 plasmids expressing viral polymerase proteins PB1, PB2, PA, NP (minigenome) and reporter construct. Results: Based on segment 8, two reporter constructs were created that contain a direct or inverted NS1-GFP-NS2 sequence for the expression of NS2 and NS1 proteins translationally fused with green fluorescent protein (GFP), which allowed the evaluation the transcriptional and/or replicative activity of viral polymerase. Conclusion: Polymerase of virus A/Krasnodar/101/35/59 (H2N2) has higher replicative and transcriptional activity at 33 °C than at 37 °C. Its transcriptional activity is more temperature-dependent than its replicative activity. The replicative and transcriptional activity of polymerase A/Puerto Rico/8/34 virus (H1N1, Mount Sinai variant) have no significant differences and do not depend on temperature.
Databáze:	MEDLINE
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