Characterization of the herpes simplex virus type 1 glycoprotein D mRNA and expression of this protein in Xenopus oocytes

Roger J. Watson, Anamaris M. Colberg-poley, Carol J. Marcus-sekura, Barrie J. Carter, Lynn W. Enquist

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

We have identified and characterized a 3.0 kilobase (kb) mRNA containing coding sequences of the herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) gene. The synthesis of this 3.0 kb mRNA was unaffected by the presence of cytosine arabinoside, but was made in greatly reduced amounts in cells infected with HSV-1 in the presence of cycloheximide: it was, therefore, classified as an early mRNA. By nuclease protection experiments, it was found that the 3.0 kb mRNA is unspliced and, further, that it is 3′ co-terminal with a smaller 1.6 kb early mRNA which is transcribed from a DNA sequence 3′ to the gD coding sequence. We describe the use of the Xenopus laevis oocyte system to produce HSV-1 gD in vitro. Oocytes injected with mRNA isolated from HSV-1-infected Vero cells synthesized gD, which was identified by immunoprecipitation. Injection of a plasmid clone containing the HSV-1 BamHI J fragment (0.89 to 0.93 map units) into the nuclei of Xenopus oocytes also resulted in synthesis of gD.

Original languageEnglish (US)
Pages (from-to)1507-1522
Number of pages16
JournalNucleic acids research
Volume11
Issue number5
DOIs
StatePublished - Mar 11 1983
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Genetics

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