Herpesviruses carrying a Brainbow cassette reveal replication and expression of limited numbers of incoming genomes.
Whether all the infectious herpesvirus particles entering a cell are able to replicate and/or express their genomes is not known. Here, we developed a general method to determine the number of viral genomes expressed in an infected cell. We constructed and analysed fluorophore expression from a recombinant pseudorabies virus (PRV263) carrying a Brainbow cassette (Cre-conditional expression of different fluorophores). Using three isogenic strains derived from PRV263, each expressing a single fluorophore, we analysed the colour composition of cells infected with these three viruses at different multiplicities. We estimate that fewer than seven incoming genomes are expressed per cell. In addition, those templates that are expressed are the genomes selected for replication and packaging into virions. This finite limit on the number of viral genomes that can be expressed is an intrinsic property of the infected cell and may be influenced by viral and cellular factors.
Department of Molecular Biology and the Princeton Neuroscience Institute, Princeton University, Princeton, New Jersey 08544, USA.
Oren
O
Kobiler
Yaron
Y
Lipman
Kate
K
Therkelsen
Ingrid
I
Daubechies
Lynn W
LW
Enquist
D000818:Animals; D002460:Cell Line; D004261:DNA Replication / Q000235:genetics*; D016679:Genome, Viral / Q000235:genetics*; D006564:Herpesviridae / Q000235:genetics*; D008856:Microscopy, Fluorescence; D008962:Models, Theoretical; D013552:Swine
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