Characterization of the Dynamic Transcriptome of a Herpesvirus with Long-read Single Molecule Real-Time Sequencing

• DOI: 10.1038/srep43751
• Publication Date: 2017-03-03T12:49:41Z
• Authors: Dóra Tombácz; Zsolt Balázs; Zsolt Csabai; Norbert Moldován; Attila Szűcs; Donald Sharon; Michael Snyder; Zsolt Boldogkői
• Keywords: Massive parallel sequencing; Lytic cycle; Illumina dye sequencing

Abstract Herpesvirus gene expression is co-ordinately regulated and sequentially ordered during productive infection. The viral genes can be classified into three distinct kinetic groups: immediate-early, early, late classes. In this study, a massively parallel sequencing technique that based on PacBio Single Molecule Real-time platform, was used for quantifying the poly(A) fraction of lytic transcriptome pseudorabies virus (PRV) throughout 12-hour interval infection PK-15 cells. Other approaches, including microarray, real-time RT-PCR Illumina are capable detecting only aggregate transcriptional activity particular genomic regions, but not individual herpesvirus transcripts. However, SMRT allows distinction between transcript isoforms, length- splice variants, as well overlapping polycistronic RNA molecules. non-amplified Isoform Sequencing (Iso-Seq) method to analyse properties PRV transcripts then classify them accordingly. Additionally, present study demonstrates general utility long-read time-course analysis global in practically any organism.