Handa H, Carter B J
J Biol Chem. 1979 Jul 25;254(14):6603-10.
A complex which is active in in vitro synthesis of adeno-associated virus (AAV) DNA was solubilized from Vero cells that were co-infected with AAV and either adenovirus (Ad5) or a herpes simplex virus type 1 (HSV-1) as the helper virus. The complexes from the Ad5 and HSV-1-infected cells sedimented at 23 S and 28 S, respectively. The optimal conditions for in vitro DNA synthesis for the two types of complex using the endogenous AAV template and the endogenous DNA polymerase, differed with respect to the effect of KCl and K2SO4 concentration. In addition the complex from HSV-1-infected cells, but not that from Ad5-infected cells, was inhibited by phosphonoacetic acid. Thus, the two complexes appear to contain different DNA polymerase activities. This was verified by phosphocellulose chromatography of the DNA polymerases solubilized from the isolated complexes. The major activity in the complex from HSV-1 infected cells was the HSV-induced DNA polymerase with lesser amounts of cellular DNA polymerase alpha and gamma or both. The complex from the Ad5-infected cells contained mainly a cellular DNA polymerase gamma.
一种在体外合成腺相关病毒（AAV）DNA中具有活性的复合物，是从同时感染了AAV以及作为辅助病毒的腺病毒（Ad5）或单纯疱疹病毒1型（HSV-1）的非洲绿猴肾细胞（Vero细胞）中溶解出来的。来自Ad5感染细胞和HSV-1感染细胞的复合物分别在23S和28S处沉降。使用内源性AAV模板和内源性DNA聚合酶，两种类型复合物进行体外DNA合成的最佳条件，在氯化钾（KCl）和硫酸钾（K2SO4）浓度的影响方面有所不同。此外，来自HSV-1感染细胞的复合物，但不是来自Ad5感染细胞的复合物，受到膦甲酸的抑制。因此，这两种复合物似乎含有不同的DNA聚合酶活性。从分离出的复合物中溶解出的DNA聚合酶经磷酸纤维素层析对此进行了验证。来自HSV-1感染细胞的复合物中的主要活性是HSV诱导的DNA聚合酶，还有少量的细胞DNA聚合酶α和γ，或者两者都有。来自Ad5感染细胞的复合物主要含有一种细胞DNA聚合酶γ。
