Silverstein S, Bachenheimer S L, Frenkel N, Roizman B
Proc Natl Acad Sci U S A. 1973 Jul;70(7):2101-4. doi: 10.1073/pnas.70.7.2101.
Analysis of the hybridization kinetics of labeled DNA of herpes simplex virus with unlabeled excess RNA from infected cells showed that viral RNA sequences form two classes differing in molar concentration. The abundant class constituted 93.5-99.3% of total virus-specific RNA and was complementary to 14-16% of the early DNA (2 hr after infection) and to 19-22% of the late DNA (8 hr after infection) in the reproductive cycle of the virus. The early RNA sequences were found to be a subset of the late sequences. The scarce sequences constituted 0.7-6.5% of total virus-specific RNA and were complementary to 28-30% of DNA both early and late in the reproductive cycle. In this study, abundant and scarce sequences were quantitatively separated on the basis of the finding that abundant species are adenylated, i.e., contain post-transcriptionally added poly(A), whereas the scarce RNA is not. Thus, nuclear and polyribosomal adenylated RNA were complementary to 24 and 22%, respectively, of viral DNA and, in abundance competition tests, were found to compete with each other and with abundant RNA from infected cells after 8 hr. The nonadenylated polyribosomal RNA was complementary to 27% of total viral DNA of which 6% was also complementary to adenylated polyribosomal RNA. Hybridization kinetics indicated that each of the fractionated adenylated RNA formed two classes complementary to 6 and 21% of viral DNA.
对单纯疱疹病毒标记DNA与感染细胞中未标记的过量RNA的杂交动力学分析表明，病毒RNA序列形成了两类摩尔浓度不同的序列。丰富类占病毒特异性RNA总量的93.5 - 99.3%，在病毒复制周期中，与早期DNA（感染后2小时）的14 - 16%以及晚期DNA（感染后8小时）的19 - 22%互补。早期RNA序列被发现是晚期序列的一个子集。稀少序列占病毒特异性RNA总量的0.7 - 6.5%，在复制周期的早期和晚期都与28 - 30%的DNA互补。在这项研究中，基于丰富序列被腺苷酸化，即含有转录后添加的多聚（A），而稀少RNA没有这一发现，对丰富和稀少序列进行了定量分离。因此，核内和多核糖体腺苷酸化RNA分别与病毒DNA的24%和22%互补，并且在丰度竞争试验中，发现它们在感染8小时后相互竞争，并与感染细胞中的丰富RNA竞争。未腺苷酸化的多核糖体RNA与病毒总DNA的27%互补，其中6%也与腺苷酸化多核糖体RNA互补。杂交动力学表明，每个分离的腺苷酸化RNA部分都形成了与病毒DNA的6%和21%互补的两类。
