Watson R J, Colberg-Poley A M, Marcus-Sekura C J, Carter B J, Enquist L W
Nucleic Acids Res. 1983 Mar 11;11(5):1507-22. doi: 10.1093/nar/11.5.1507.
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.
我们已经鉴定并描述了一种含有单纯疱疹病毒1型（HSV-1）糖蛋白D（gD）基因编码序列的3.0千碱基（kb）信使核糖核酸（mRNA）。这种3.0 kb mRNA的合成不受阿糖胞苷存在的影响，但在存在环己酰亚胺的情况下感染HSV-1的细胞中其合成量大幅减少：因此，它被归类为早期mRNA。通过核酸酶保护实验发现，3.0 kb mRNA未经过剪接，而且，它与一个较小的1.6 kb早期mRNA在3'端共末端，该1.6 kb早期mRNA从gD编码序列下游的一个DNA序列转录而来。我们描述了使用非洲爪蟾卵母细胞系统在体外产生HSV-1 gD的方法。注射从感染HSV-1的非洲绿猴肾细胞（Vero细胞）中分离出的mRNA的卵母细胞合成了gD，通过免疫沉淀法得以鉴定。将含有HSV-1 BamHI J片段（0.89至0.93个图谱单位）的质粒克隆注射到非洲爪蟾卵母细胞核中也导致了gD的合成。
