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Nucleic Acids Research, 1983, Vol. 11, No. 5 1507-1522
© 1983

MOLECULAR BIOLOGY

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

Roger J. Watson1, Anamaris M. Colberg-Poley2, Carol J. Marcus-Sekura2, Barrie J. Carter2 and Lynn W. Enquist1

1Molecular Genetics, Inc. 10320 Bren Road East, Minnetonka, MN 55343 2laboratory of Cell Biology and Genetics, NIADDK Building 4, Room 312 National Institutes of Health Bethesda, MD 20205, USA

Received October 8, 1982. Revised February 1, 1983. Accepted February 1, 1983.

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.


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P. Berman, D Dowbenko, L. Lasky, and C. Simonsen
Detection of antibodies to herpes simplex virus with a continuous cell line expressing cloned glycoprotein D
Science, November 4, 1983; 222(4623): 524 - 527.
[Abstract] [PDF]


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