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Population Variability Generated during Rescue Process and Passaging of Recombinant Mumps Viruses

Slović, Anamarija; Košutić Gulija, Tanja; Forčić, Dubravko; Šantak, Maja; Jagušić, Maja; Jurković, Mirna; Pali, Dorotea; Ivančić-Jelečki, Jelena (2021) Population Variability Generated during Rescue Process and Passaging of Recombinant Mumps Viruses. Viruses-Basel, 13 (12). ISSN 1999-4915

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Recombinant mumps viruses (MuVs) based on established vaccine strains represent attractive vector candidates as they have known track records for high efficacy and the viral genome does not integrate in the host cells. We developed a rescue system based on the consensus sequence of the L- Zagreb vaccine and generated seven different recombinant MuVs by (a) insertion of one or two additional transcription units (ATUs), (b) lengthening of a noncoding region to the extent that the longest noncoding region in MuV genome is created, or (c) replacement of original L-Zagreb sequences with sequences rich in CG and AT dinucleotides. All viruses were successfully rescued and faithfully matched sequences of input plasmids. In primary rescued stocks, low percentages of heterogeneous positions were found (maximum 0.12%) and substitutions were predominantly obtained in minor variants, with maximally four substitutions seen in consensus. ATUs did not accumulate more mutations than the natural MuV genes. Six substitutions characteristic for recombinant viruses generated in our system were defined, as they repetitively occurred during rescue processes. In subsequent passaging of primary rescue stocks in Vero cells, different inconsistencies within quasispecies structures were observed. In order to assure that unwanted mutations did not emerge and accumulate, sub-consensus variability should be closely monitored. As we show for Pro408Leu mutation in L gene and a stop codon in one of ATUs, positively selected variants can rise to frequencies over 85% in only few passages.

Item Type: Article
Uncontrolled Keywords: virus rescue ; mumps virus ; virus variability
Subjects: NATURAL SCIENCES > Biology
Divisions: Division of Physical Chemistry
Project titleProject leaderProject codeProject type
Centar izvrsnosti za virusnu imunologiju i cjepiva-CERVirVacStipan JonjićN/AMZOS
Depositing User: Kristina Ciglar
Date Deposited: 07 Apr 2022 14:09
DOI: 10.3390/v13122550

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