Genotipizacija virusa Epstein-Barr analizom sekvencije gena LMP-1

master's thesis

• Title (english): Epstein-Barr virus genotyping based on LMP-1 gene sequencing
• Author: Margarita Batović
• Mentor: Snježana Židovec Lepej (mentor)
• Committee member: Snježana Židovec Lepej (predsjednik povjerenstva)
• Committee member: Silvija Černi (član povjerenstva)
• Committee member: Nada Oršolić (član povjerenstva)
• Committee member: Petra Korać (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2019-03-19, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Virus Epstein-Barr (EBV) ili humani gammaherpesvirus 4 je DNA virus koji pripada rodu Lymphocryptovirus. EBV se klasificira u podtipove prema genetskoj raznolikosti gena LMP-1 koji kodira latentni membranski protein virusa te pokazuje viši stupanj polimorfizma u usporedbi s drugim genima ovog virusa. Cilj ovog rada bio je odrediti distribuciju podtipova virusa EBV temeljem sekvencije karboksi-terminalne regije gena LMP-1. U istraživanje je uključen 41 ispitanik s infektivnom mononukleozom uzrokovanom infekcijom virusom EBV. Podtipovi virusa EBV određeni su metodom Sangerovog sekvenciranja i filogenetske analize. Rezultati ovog istraživanja po prvi su puta dokazali prisutnost podtipova China 1, Mediterranean, B95-8 i North Carolina virusa EBV koji se temelje na sekvenci karboksi-terminalne regije gena LMP-1. Analizom sekvencija LMP-1 gena dokazana je prisutnost delecija veličine 30 pb i 69 pb, kao i karakteristična tandemska ponavljanja veličine 33 pb. Detektirano je čak 60 aminokiselinskih promjena pojedinih LMP-1 podtipova virusa u odnosu na EBV prototip B95-8. U ovom su radu otkrivene četiri nove supstitucije u izolatima podtipa Mediterranean, kao i u prototipu B95-8 virusa. Dokazana je i povezanost kinetike virusne replikacije i EBV LMP-1 podtipova kao i negativna korelacija između dobi ispitanika i viremije. Rezultati ovog rada ukazuju na molekularnu raznolikost virusa Epstein-Barr u Hrvatskoj.

Abstract (english)

Epstein-Barr virus (EBV) or human gammaherpes virus 4 is a DNA virus belonging to the genus Lymphocryptovirus. Classification of EBV subtypes is based on genetic diversity of LMP-1 gene which is coding for the synthesis of latent membrane protein 1 and exhibits a higher degree of polymorphism in comparison to the other EBV genes. The main goal of this study was to determine the distribution of EBV subtypes based on the sequences of LMP-1 gene carboxy-terminal region. This study included 41 participants with infectious mononucleosis caused by EBV infection. EBV subtypes were determined by Sanger's method of sequencing and phylogenetic analysis. Results of this research showed, for the first time, the presence of China1, Mediterranean, B95-8 and North Carolina subtypes of EBV based on a carboxy-terminal LMP-1 gene sequence, while LMP-1 gene analysis showed the presence of 30 bp and 69 bp deletions and characteristic 33 bp tandem repeats. A total of 60 amino acid changes in individual viral LMP-1 subtypes compared to the B95-8 prototype strain were detected. Four new substitutions were discovered within Mediterranean and B95-8 subtypes. Furthermore, association between viral replication kinetics and EBV LMP-1 subtypes was also found. The results of this study indicate a molecular diversity of Epstein-Barr virus in Croatia.

• Keywords: infektivna mononukleoza; podgenotipovi; molekularna filogenija; sekvenciranje
• Keywords (english): infectious mononucleosis; subgenotypes; molecular phylogeny; sequencing
• Language: croatian
• URN:NBN: urn:nbn:hr:217:294826
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2021-03-19
• Created on: 2019-04-10 09:28:01