Lee L F, Boezi J A, Blakesley R W, Koenig M, Towle H C
J Virol. 1974 Nov;14(5):1209-19. doi: 10.1128/JVI.14.5.1209-1219.1974.
Infection of duck embryo fibroblasts by Marek's disease herpesvirus (MDHV), strain GA, led to the induction of a novel DNA polymerase. This novel DNA polymerase, designated MDHV-induced DNA polymerase, could be distinguished from the DNA polymerase activities of uninfected duck embryo fibroblasts by its chromatographic behavior on phosphocellulose, by its sedimentation coefficient, and by its catalytic properties. The characteristics of MDHV-induced DNA polymerase which had been purified by phosphocellulose chromatography were investigated. The sedimentation coefficient of the enzyme, as determined by sucrose density gradient centrifugation in the presence of 0.25 M KCl, was 5.9S. From this sedimentation coefficient, the molecular weight of MDHV-induced DNA polymerase was estimated to be 100,000. MDHV-induced DNA polymerase could not effectively use either poly(dA).oligo(dT)(12-18) or poly(dC).oligo(dG)(12-18) as a template-primer. The DNA polymerases from uninfected duck embryo fibroblasts could use these synthetic template-primers. MDHV-induced DNA polymerase also could not use poly(rA).oligo(dT)(12-18) or poly(rC).oligo(dG)(12-18) as template-primers or oligo(dT)(12-18) as a primer, indicating that it was not a polymerase of the type R-DNA polymerase, reverse transcriptase, or a terminal nucleotidyl transferase. In vitro synthesis of DNA by MDHV-induced DNA polymerase was markedly inhibited by the addition of (NH(4))(2)SO(4) to the reaction mixture.
马立克氏病疱疹病毒(MDHV)GA株感染鸭胚成纤维细胞后,诱导产生了一种新型DNA聚合酶。这种新型DNA聚合酶被命名为MDHV诱导的DNA聚合酶,通过其在磷酸纤维素上的色谱行为、沉降系数及其催化特性,可与未感染鸭胚成纤维细胞的DNA聚合酶活性区分开来。对通过磷酸纤维素色谱法纯化的MDHV诱导的DNA聚合酶的特性进行了研究。在0.25M KCl存在下,通过蔗糖密度梯度离心法测定该酶的沉降系数为5.9S。根据这个沉降系数,估计MDHV诱导的DNA聚合酶的分子量为100,000。MDHV诱导的DNA聚合酶不能有效地将聚(dA)·寡聚(dT)(12 - 18)或聚(dC)·寡聚(dG)(12 - 18)用作模板引物。未感染鸭胚成纤维细胞的DNA聚合酶可以使用这些合成模板引物。MDHV诱导的DNA聚合酶也不能将聚(rA)·寡聚(dT)(12 - 18)或聚(rC)·寡聚(dG)(12 - 18)用作模板引物,也不能将寡聚(dT)(12 - 18)用作引物,这表明它不是R型DNA聚合酶、逆转录酶或末端核苷酸转移酶类型的聚合酶。向反应混合物中添加硫酸铵会显著抑制MDHV诱导的DNA聚合酶在体外合成DNA。