Sarin P S, Donlon J, Friedman B, Gallo R C
Biochim Biophys Acta. 1979 Sep 27;564(2):235-45. doi: 10.1016/0005-2787(79)90222-3.
An RNA-directed DNA polymerase was purified from a cell line derived from a radiation-induced lymphoma in NIH Swiss mice which produced non-infectious type C virus particles. The enzyme was isolated from a high speed particulate fraction which bands at a density of 1.16--1.19 g/ml in a sucrose gradient, and purified by successive chromatography on DEAE-cellulose, phosphocellulose and hydroxyapatite. The purified DNA polymerase has a molecular weight of 68 000, a pH optimum of 7.5, a KCl optimum of 50 mM, and a Mn2+ optimum of 0.25 mM. It prefers (dT)15 . (A)n to (dT)15 . (dA)n as the primer template and transcribes the poly(C) strand of (dG)15 .(C)n and (dG)15 . (OMeC)n. It transcribes heteropolymeric regions of avian myeloblastosis virus 70 S RNA, and is inhibited by antiserum to Rauscher murine leukemia virus DNA polymerase. Comparison of the properties of DNA polymerase purified from radiation-induced lymphoma cells with the DNA polymerase purified from non-defective murine type C RNA tumor viruses shows that the mouse lymphoma enzyme is both biochemically and immunologically related to murine leukemia virus DNA polymerases.
从NIH瑞士小鼠辐射诱导的淋巴瘤细胞系中纯化出一种RNA指导的DNA聚合酶，该细胞系可产生非感染性C型病毒颗粒。该酶从高速颗粒组分中分离得到，在蔗糖梯度中密度为1.16 - 1.19 g/ml处形成条带，并通过在DEAE - 纤维素、磷酸纤维素和羟基磷灰石上连续层析进行纯化。纯化后的DNA聚合酶分子量为68000，最适pH为7.5，最适KCl浓度为50 mM，最适Mn2+浓度为0.25 mM。它更倾向于以(dT)15.(A)n而非(dT)15.(dA)n作为引物模板，并转录(dG)15.(C)n和(dG)15.(OMeC)n的聚(C)链。它能转录禽成髓细胞瘤病毒70 S RNA的杂聚区域，并被劳舍尔鼠白血病病毒DNA聚合酶抗血清所抑制。将从辐射诱导的淋巴瘤细胞中纯化的DNA聚合酶的性质与从无缺陷的鼠C型RNA肿瘤病毒中纯化的DNA聚合酶的性质进行比较，结果表明小鼠淋巴瘤酶在生化和免疫方面均与鼠白血病病毒DNA聚合酶相关。
