Naslov
Resekvenciranje genoma virusa Puumala iz WHO arbovirusne kolekcije
(Resequencing of the Puumala virus strain Sotkamo from the WHO Arbovirus collection)

Autori
Kurolt, Ivan-Christian ; Paessler, Slobodan ; Markotić, Alemka

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
CROCMID 2013 Knjiga sažetaka/Abstract book / Bradarić, Nikola ; Tambić Andrašević, Arjana - Zagreb : Hrvatski liječnički zbor ; Hrvatsko društvo za mikrobiologiju ; Hrvatsko društvo za infektivne bolesti, 2013, 204-205

Skup
10. hrvatski kongres kliničke mikrobiologije i 7. hrvatski kongres o infektivnim bolestima

Mjesto i datum
Rovinj, Hrvatska, 24.10.2013. - 27.10.2013

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
PUUV; genomska sekvenca; resekvenciranje; SZO arbovirusna kolekcija
(PUUV; Genome sequence; Resequencing; WHO Arbovirus collection)

Sažetak
The inability of RNA polymerase to perform proofreading during replication causes high mutation rates in Hantaviruses and other RNA viruses. Our goal was to clone and determine the sequence of the Puumala virus strain Sotkamo from WHO arbovirus collection (Dr. R. Tesh, UTMB) and to compare it to the published sequence. RNA from VeroE6 cells infected with Puumala virus strain Sotkamo was isolated using chloroform-phenol extraction and the cDNA was prepared using random hexamers. Viral sequences were amplified using specific primers and cloned into suitable vectors. At least three clones of each fragment were sequenced using dye termination sequencing on Applied Biosystems Model 3100 Genetic Analyzer. The sequence obtained for the three segments showed more than 99% homology compared to the published genome for the Puumala Sotkamo strain. Sequencing showed seven mutations in the S segment, four in the non-coding region and three silent mutations in the open reading frame for the nucleocapsid protein. M segment contained eleven mutations in the coding region for Puumala glycoprotein, all but two caused amino acid changes. Only five mutations were observed in L segment, causing four amino acid changes in the 6471 nucleotides long open reading frame for the viral polymerase. Interestingly, these mutations aren't spread evenly through the genome. The L segment shows proportionally the smallest number of mutations, whereas M and S segments have a higher but proportionally similar amount of changes. However, their potential effects differ within the genome segment. While 80% of mutations cause an amino acid change in the M segment, all mutations are silent in the S segment. The segment's non-coding region probably accumulates more nucleotide changes than the coding region.

Izvorni jezik
Hrvatski, engleski

Znanstvena područja
Biologija, Temeljne medicinske znanosti

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