title: Visualization of herpes simplex virus type 1 virions using fluorescent colors.
abstract: Our laboratory was one of the first to engineer a live fluorescent tag, enhanced green fluorescent protein (eGFP), that marked the capsid of herpes simplex virus type 1 (HSV-1) and subsequently maturing virus as the particle made its way to the cell surface. In the present study we sought to increase the repertoire of colors available as fusion to the small capsid protein, VP26, so that they can be used alone or in conjunction with other fluorescent tags (fused to other HSV proteins) to follow the virus as it enters and replicates within the cell. We have now generated viruses expressing VP26 fusions with Cerulean, Venus, mOrange, tdTomato, mCherry, and Dronpa3 fluorescent proteins. These fusions were made in a repaired UL35 gene (VP26) background. These fusions do not affect the replication properties of the virus expressing the fusion polypeptide and the fusion tag was stably associated with intranuclear capsids and mature virions. Of note we could not isolate viruses expressing fusions with fluorescent proteins that have a tendency to dimerize.
authors.affiliation: Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Lyns
authors.initials: L
authors.lastname: Etienne
authors.affiliation: Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Poorval
authors.initials: P
authors.lastname: Joshi
authors.affiliation: Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Dingle
authors.affiliation: Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Eugene
authors.initials: E
authors.lastname: Huang
authors.affiliation: Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Grzesik
authors.affiliation: Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, The Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: pdesai@jhmi.edu.
authors.forename: Prashant J
authors.initials: PJ
authors.lastname: Desai
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000235:genetics; D002522:Chlorocebus aethiops; D003116:Color; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D014709:Vero Cells; D014779:Virus Replication; D000097573:Red Fluorescent Protein
chemical_list: D036022:Capsid Proteins; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein; C077409:capsid protein VP26, herpes simplex virus type 1; C485184:enhanced green fluorescent protein; C413663:fluorescent protein 583
keywords: Capsid; Fluorescent tags; HSV1; VP26; Venus; tdTomato