Whole-genome analysis of pseudorabies virus gene expression by real-time quantitative RT-PCR assay

BMC Genomics
Dóra TombáczZsolt Boldogköi


Pseudorabies virus (PRV), a neurotropic herpesvirus of pigs, serves as an excellent model system with which to investigate the herpesvirus life cycle both in cultured cells and in vivo. Real-time RT-PCR is a very sensitive, accurate and reproducible technique that can be used to detect very small amounts of RNA molecules, and it can therefore be applied for analysis of the expression of herpesvirus genes from the very early period of infection. In this study, we have developed and applied a quantitative reverse transcriptase-based real-time PCR technique in order to profile transcription from the whole genome of PRV after lytic infection in porcine kidney cells. We calculated the relative expression ratios in a novel way, which allowed us to compare different PRV genes with respect to their expression dynamics, and to divide the PRV genes into distinct kinetic classes. This is the first publication on the whole-genome analysis of the gene expression of an alpha-herpesvirus by qRT2-PCR. We additionally established the kinetic properties of uncharacterized PRV genes and revised or confirmed data on PRV genes earlier examined by traditional methods such as Northern blot analysis. Our investigations revealed that genes with the sam...Continue Reading


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Methods Mentioned


Related Concepts

DNA Replication
Genes, Reiterated
Herpesvirus 1, Suid
Protein Biosynthesis
Gene Expression Regulation, Viral
Viral Genome
Reverse Transcriptase Polymerase Chain Reaction
MRNA Differential Display

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