藜（Chenopodium album L.）中一种γ样DNA聚合酶的纯化与特性分析

Purification and characterization of a gamma-like DNA polymerase from Chenopodium album L.

作者信息

Meissner K, Heinhorst S, Cannon G C, Börner T

机构信息

FB Biologie, Humboldt-Universität Berlin, Germany.

出版信息

Nucleic Acids Res. 1993 Oct 25;21(21):4893-9. doi: 10.1093/nar/21.21.4893.

DOI:10.1093/nar/21.21.4893

PMID:8177736

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC311402/

Abstract

A DNA polymerase activity from mitochondria of the dicotyledonous angiosperm Chenopodium album L. was purified almost 9000 fold by successive column chromatography steps on DEAE cellulose, heparin agarose and ssDNA cellulose. The enzyme was characterized as a gamma-class polymerase, based on its resistance to inhibitors of the nuclear DNA polymerase alpha and its preference for poly(rA).(dT)12-18 over activated DNA in vitro. The molecular weight was estimated to be 80,000-90,000. A 3' to 5' exonuclease activity was found to be tightly associated with the DNA polymerase activity through all purification steps. This is the first report of an association between a DNA polymerase and an exonuclease activity in plant mitochondria.

摘要

通过在DEAE纤维素、肝素琼脂糖和单链DNA纤维素上连续进行柱层析步骤，将双子叶被子植物藜（Chenopodium album L.）线粒体中的一种DNA聚合酶活性纯化了近9000倍。基于其对核DNA聚合酶α抑制剂的抗性以及在体外对聚（rA）·（dT）12 - 18而非活化DNA的偏好，该酶被鉴定为γ类聚合酶。估计其分子量为80,000 - 90,000。在所有纯化步骤中，发现3'至5'核酸外切酶活性与DNA聚合酶活性紧密相关。这是关于植物线粒体中DNA聚合酶与核酸外切酶活性之间关联的首次报道。