Yanagi K, Rush M G, Biegeleisen K
J Gen Virol. 1979 Sep;44(3):657-67. doi: 10.1099/0022-1317-44-3-657.
The ability of HSV-1 DNA to become associated with host cell DNA in an alkaline-stable form has been demonstrated following infection of a ts baby hamster kidney growth mutant (ts BTN-1), at the non-permissive temperature (39.5 depgrees C). After 8 h pre-incubation at 39.5 degrees C, ts BTN-1 cells infected at this temperature using m.o.i. ranging from 0.5 to 200 p.f.u./cell fail to replicate virus DNA even though transport of input virus genomes to the nucleus is the same at both permissive and non-permissive temperatures. Virions containing 3H-labelled DNA were used to infect ts BTN-1 cells at 39.5 degrees C, and the total cellular DNA isolated from these cells was resolved into host and virus material by repeated CsCl equilibrium gradient centrifugation. A significant amount of the input radioactivity was found as a distinct band in the host region in both neutral and alkaline CsCl gradients, strongly suggesting a covalent association between host and virus DNAs. Evidence for this association was strengthened by demonstrating that radioactive material (virus DNA) banding in the host region of CsCl gradients could be driven towards the density expected for virus DNA following degradation of the putative hybrid molecules by shearing.
在非允许温度（39.5摄氏度）下，对温度敏感的幼仓鼠肾生长突变体（ts BTN-1）进行感染后，已证明单纯疱疹病毒1型（HSV-1）DNA能够以碱稳定形式与宿主细胞DNA结合。在39.5摄氏度下预孵育8小时后，在此温度下使用感染复数（m.o.i.）为0.5至200个噬斑形成单位/细胞感染的ts BTN-1细胞无法复制病毒DNA，尽管在允许温度和非允许温度下输入病毒基因组向细胞核的转运是相同的。用含有3H标记DNA的病毒粒子在39.5摄氏度下感染ts BTN-1细胞，通过反复的CsCl平衡梯度离心将从这些细胞中分离的总细胞DNA分离为主细胞和病毒物质。在中性和碱性CsCl梯度中，在宿主区域均发现大量输入放射性作为一条明显的条带，强烈表明宿主和病毒DNA之间存在共价结合。通过证明在假定的杂交分子被剪切降解后，CsCl梯度宿主区域中放射性物质（病毒DNA）条带可朝着病毒DNA预期的密度移动，进一步加强了这种结合的证据。
