Cloning and sequencing of Puumala virus Sotkamo strain S and M RNA segments: evidence for strain variation in hantaviruses and expression of the nucleocapsid protein
| Autor: | Antti Vaheri, Hannimari Kallio-Kokko, Olli Vapalahti, Eeva-Marjatta Salonen, M. Brummer-Korvenkontio |
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| Rok vydání: | 1992 |
| Předmět: |
Orthohantavirus
Reading Frames Recombinant Fusion Proteins Blotting Western Molecular Sequence Data Sequence alignment Molecular cloning Polymerase Chain Reaction Viral Proteins 03 medical and health sciences Sequence Homology Nucleic Acid Virology Animals Amino Acid Sequence Cloning Molecular Vero Cells 030304 developmental biology Genomic organization chemistry.chemical_classification 0303 health sciences Base Sequence biology 030306 microbiology Nucleic acid sequence Genetic Variation RNA beta-Galactosidase biology.organism_classification Molecular biology Amino acid Open reading frame chemistry Hemorrhagic Fever with Renal Syndrome RNA Viral Puumala virus |
| Zdroj: | Journal of General Virology. 73:829-838 |
| ISSN: | 1465-2099 0022-1317 |
| DOI: | 10.1099/0022-1317-73-4-829 |
| Popis: | The prototype Puumala virus (PV) Sotkamo strain small (S) and medium (M) RNA genome segments were amplified by the polymerase chain reaction (PCR), cloned and sequenced. The S segment is 1830 nucleotides long with an open reading frame coding for 433 amino acids. The identity to the PV Hällnäs strain was 83% at the nucleotide and 96% at the amino acid level. The M segment in the Sotkamo strain is 3616 nucleotides long and contains one open reading frame of 1148 amino acids with 83% nucleotide and 94% amino acid identity to the Hällnäs strain. Most amino acid changes were conservative and the five predicted glycosylation sites are identical. The amino acid identity to the prototype hantavirus, Hantaan virus, was 62 and 54% for S and M segments, respectively. The coding region of the S segment was further amplified by PCR, ligated to pEX vectors and expressed in Escherichia coli as a beta-galactosidase fusion protein and was seen to be specifically detected by nephropathia epidemica sera in immunoblotting. |
| Databáze: | OpenAIRE |
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