从感染劳斯氏白血病病毒的小鼠脾脏中纯化RNA指导的DNA聚合酶。

Purification of RNA-directed DNA polymerase from mouse spleen infected with Rauscher leukemia virus.

作者信息

Drescher B, Riedel H, Niemiec P

出版信息

Acta Biol Med Ger. 1978;37(11-12):1655-63.

Abstract

RNA-directed DNA polymerase was purified from spleens of Balb/c and NMRI mice infected with Rauscher murine leukemia virus. The method includes cell fractionation and lysis of microsomal fraction, chromtography on Sephadex G-200 and phosphocellulose. Estimation of molecular weight from the sedimentation rate of the purified enzyme in a glycerol gradient was consistent with a structure containing one polypeptide with a molecular weight of 70,000. Purified RLV DNA polymerase from spleen could transcribe purified DNA polymerase from purified virions. This simple preparation method offers a procedure for large scale preparation of the RNA-directed DNA polymerase which can be used for synthesis of DNA complementary to mRNA.

摘要

从感染劳斯氏鼠白血病病毒的Balb/c和NMRI小鼠脾脏中纯化出RNA指导的DNA聚合酶。该方法包括细胞分级分离和微粒体部分的裂解、在葡聚糖G-200和磷酸纤维素上的色谱法。根据纯化酶在甘油梯度中的沉降速率估算分子量，结果与含有一条分子量为70,000的多肽的结构一致。从脾脏中纯化的劳斯氏鼠白血病病毒DNA聚合酶可以转录来自纯化病毒粒子的纯化DNA聚合酶。这种简单的制备方法为大规模制备RNA指导的DNA聚合酶提供了一种程序，该酶可用于合成与mRNA互补的DNA。