Nonstructural proteins of herpes simplex virus. I. Purification of the induced DNA polymerase
Nonstructural proteins of herpes simplex virus. I. Purification of the induced DNA polymerase
复制标题
单纯疱疹病毒的非结构蛋白。
作者：
K. Powell;D. Purifoy
用已发表的方法纯化的单纯疱疹病毒诱导的DNA聚合酶被发现被许多其他蛋白质污染，包括病毒结构蛋白。因此，DEAE-纤维素和磷酸纤维素层析与亲和层析结合使用，以从单纯疱疹病毒1型和2型感染的细胞中纯化DNA聚合酶。纯化的酶保留了疱疹病毒诱导的DNA聚合酶的独特功能，包括最大活性需要高盐浓度，对低膦酰乙酸浓度的敏感性，以及被疱疹病毒感染细胞的兔抗血清中和的能力。通过聚丙烯酰胺凝胶电泳，纯化的DNA聚合酶与约150，000分子量的病毒诱导的多肽结合。
Herpes simplex virus-induced DNA polymerase purified by published methods was found to be contaminated with many others proteins, including virus structural proteins. Thus, DEAE-cellulose and phosphocellulose chromatography were used in combination with affinity chromatography to purify DNA polymerase from herpes simplex virus type 1- and type 2-infected cells. The purified enzyme retained unique features of the herpesvirus-induced DNA polymerase, including a requirement for high salt concentrations for maximal activity, a sensitivity to low phosphonoacetate concentrations, and the capacity to be neutralized by rabbit antiserum to herpesvirus-infected cells. By polyacrylamide gel electrophoresis, the purified DNA polymerase was associated with a virus-induced polypeptide of about 150,000 molecular weight.