Lee G T, Para M F, Spear P G
J Virol. 1982 Jul;43(1):41-9. doi: 10.1128/JVI.43.1.41-49.1982.
To map the structural genes for the gD and gE polypeptides and for other viral products encoded in the S component of herpes simplex virus type 1 DNA, we selected mRNAs capable of hybridizing to cloned viral DNA fragments and translated the mRNAs in vitro to determine which polypeptides were encoded therein. The gD and gE polypeptides were identified by immunoprecipitation with appropriate monoclonal and monospecific antibodies, whereas the other polypeptides were characterized only by their electrophoretic mobilities in polyacrylamide gels. We found that gD mRNA hybridized to a single SacI subfragment of BamHI fragment J, whereas gE mRNA hybridized to an adjacent SacI subfragment of BamHI fragment J and also to BamHI fragment X. These and other results permit the conclusion that the structural gene for gD is located between map coordinates 0.911 and 0.924, and the gene for gE is between map coordinates 0.924 and 0.951. We also found that mRNAs for polypeptides of 55,000, 42,000, 33,000, and 22,000 molecular weight hybridized to DNA fragments spanning the regions from map coordinates 0.911 to 0.924, 0.897 to 0.911, 0.939 to 0.965, and 0.939 to 0.965, respectively. Finally, in accord with the results of others, we found that mRNA for a 68,000-molecular-weight polypeptide hybridized to the two noncontiguous BamHI fragments N and Z, which share a reiterated DNA sequence.
为了绘制单纯疱疹病毒1型DNA的S成分中编码gD和gE多肽以及其他病毒产物的结构基因图谱，我们选择了能够与克隆的病毒DNA片段杂交的mRNA，并在体外翻译这些mRNA，以确定其中编码了哪些多肽。通过用适当的单克隆和单特异性抗体进行免疫沉淀来鉴定gD和gE多肽，而其他多肽仅通过它们在聚丙烯酰胺凝胶中的电泳迁移率来表征。我们发现gD mRNA与BamHI片段J的单个SacI亚片段杂交，而gE mRNA与BamHI片段J的相邻SacI亚片段杂交，也与BamHI片段X杂交。这些以及其他结果使得我们得出结论，gD的结构基因位于图谱坐标0.911和0.924之间，而gE的基因位于图谱坐标0.924和0.951之间。我们还发现，分子量为55,000、42,000、33,000和22,000的多肽的mRNA分别与跨越图谱坐标0.911至0.924、0.897至0.911、0.939至0.965和0.939至0.965区域的DNA片段杂交。最后，与其他人的结果一致，我们发现分子量为68,000的多肽的mRNA与两个不连续的BamHI片段N和Z杂交，这两个片段共享一个重复的DNA序列。
