Determination of cold-adapted influenza virus (Orthomyxoviridae: Alphainfluenzavirus) polymerase activity by the minigenome method with a fluorescent protein
| Autor: | Pavel A. Ivanov, Aleksandr V. Lyashko, Vladimir Y. Kost, Natalia F. Lomakina, Artyom A. Rtishchev, Nataliya I. Bunkova, Tatiana A. Timofeeva, Marina A. Balanova, Stepan A. Ionov, Dmitry V. Gorikov, Stanislav G. Markushin |
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| Jazyk: | English<br />Russian |
| Rok vydání: | 2023 |
| Předmět: | |
| Zdroj: | Вопросы вирусологии, Vol 68, Iss 6, Pp 526-535 (2023) |
| Druh dokumentu: | article |
| ISSN: | 0507-4088 2411-2097 |
| DOI: | 10.36233/0507-4088-203 |
| Popis: | 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: | Directory of Open Access Journals |
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