单纯疱疹病毒在感染细胞的部分纯化可溶性提取物中的DNA合成。
Herpes simplex virus DNA synthesis in a partially purified soluble extract from infected cells.
作者信息
Pignatti P F, Cassai E, Bertazzoni U
出版信息
J Virol. 1979 Dec;32(3):1033-6. doi: 10.1128/JVI.32.3.1033-1036.1979.
Abstract
HEp-2 cells were infected with herpes simplex virus type 1 and extracted with 0.25% Triton X-100 and 0.1 M NaCl. The extract was sedimented on sucrose gradients, and the fractions containing the endogenous DNA polymerizing activity (replication complex) were collected. The properties of the replication complex were partially characterized. Under optimal conditions 375 pmol of dTMP per micrograms of DNA was incorporated, which corresponds to about 50% replication of preexisting viral DNA. The replication complex was shown to contain only DNA of viral origin by its density in CsCl. By using specific assays for DNA polymerases alpha, beta, gamma, and herpes simplex virus, we found that only the viral DNA polymerase copurified with the replication complex.
摘要
将1型单纯疱疹病毒感染HEp-2细胞，并用0.25% Triton X-100和0.1 M NaCl进行提取。提取物在蔗糖梯度上进行沉降，收集含有内源性DNA聚合活性（复制复合物）的级分。对复制复合物的特性进行了部分表征。在最佳条件下，每微克DNA掺入375 pmol的dTMP，这相当于约50%的原有病毒DNA复制。通过CsCl密度分析表明，复制复合物仅含有病毒来源的DNA。通过使用针对DNA聚合酶α、β、γ和单纯疱疹病毒的特异性检测方法，我们发现只有病毒DNA聚合酶与复制复合物共纯化。
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