从劳斯肉瘤病毒转化的大鼠细胞系中对DNA聚合酶进行部分纯化及特性鉴定。

Partial purification and characterization of DNA polymerases from a Rous sarcoma virus-transformed rat cell line.

作者信息

Bandyopadhyay A K

出版信息

Biochim Biophys Acta. 1975 Sep 12;407(1):1-13. doi: 10.1016/0005-2787(75)90019-2.

Abstract

A new DNA polymerase was partially purified from cell-free extracts of a continuous rat cell-line (XC). The XC cells had been transformed by the Prague strain of Rous sarcoma virus but did not produce infectious virus. The molecular weight of the DNA polymerase is 70,000, as estimated by glycerol gradient centrifugation and by Sephadex gel filtration. This enzyme can be distinguished from the other cellular DNA polymerases by its elution pattern on DNA-cellulose column chromatography, its molecular weight, and its primer-template specificity. The enzyme has some characteristics of the murine leukemia virus reverse transcriptase. It is partially inhibited by immunoglobulin G purified from rabbit antiserum prepared against Rauscher leukemia virus reverse transcriptase, but is not inhibited by IgG from rat antiserum prepared against avian myeloblastosis virus reverse transcriptase. However, the XC cell enzyme can be distinguished from the murine leukemia virus reverse transcriptase by its inefficiency in copying an oligo(dG)12-poly(rC)primer-template.

摘要

从大鼠连续细胞系（XC）的无细胞提取物中部分纯化出一种新的DNA聚合酶。XC细胞已被劳氏肉瘤病毒布拉格株转化，但不产生传染性病毒。通过甘油梯度离心和葡聚糖凝胶过滤估计，该DNA聚合酶的分子量为70,000。这种酶可以通过其在DNA - 纤维素柱色谱上的洗脱模式、分子量及其引物 - 模板特异性与其他细胞DNA聚合酶区分开来。该酶具有鼠白血病病毒逆转录酶的一些特征。它被从针对劳斯氏白血病病毒逆转录酶制备的兔抗血清中纯化的免疫球蛋白G部分抑制，但不被针对禽成髓细胞瘤病毒逆转录酶制备的大鼠抗血清中的IgG抑制。然而，XC细胞酶可以通过其在复制寡聚（dG）12 - 聚（rC）引物 - 模板方面的低效性与鼠白血病病毒逆转录酶区分开来。