TY - JOUR
T1 - Characterization of the herpes simplex virus type 1 glycoprotein D mRNA and expression of this protein in Xenopus oocytes
AU - Watson, Roger J.
AU - Colberg-poley, Anamaris M.
AU - Marcus-sekura, Carol J.
AU - Carter, Barrie J.
AU - Enquist, Lynn W.
N1 - Funding Information:
ACKNOWLEDGEMENTS ACP was a recipien t of PHS Grant Number 5 F32 CA 06913 awarded by th e National Cancer Institute, DHHS. Wethank Michael Hitchcock for instruction on the use of the Hamilton microinjection apparatus, Joan Mok for typing of the manuscript, and Martin Zweig of the National Cancer Institute (Frederick Cancer Research Center, Maryland) for his generous gift of the HSV-1 moroclonals used in this study.
PY - 1983/3/11
Y1 - 1983/3/11
N2 - 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.
AB - 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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U2 - 10.1093/nar/11.5.1507
DO - 10.1093/nar/11.5.1507
M3 - Article
C2 - 6298745
AN - SCOPUS:0021100691
SN - 0305-1048
VL - 11
SP - 1507
EP - 1522
JO - Nucleic acids research
JF - Nucleic acids research
IS - 5
ER -