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从人白血病脾脏中纯化RNA指导的DNA聚合酶。

Purification of RNA-instructed DNA polymerase from human leukemic spleens.

作者信息

Witkin S S, Ohno T, Spiegelman S

出版信息

Proc Natl Acad Sci U S A. 1975 Oct;72(10):4133-6. doi: 10.1073/pnas.72.10.4133.

Abstract

Particles possessing a density of 1.16 g/ml and encapsulating a 70S RNA and a RNA-instructed DNA polymerase (reverse transcriptase) have been prepared from the spleen of a patient with chronic lymphocytic leukemia. These particles have been converted to cores with a density of 1.26 g/ml and containing the enzyme-RNA complex, in complete analogy to the known RNA tumor viruses of avian and murine origin. The reverse transcriptase was purified from the cores by column chromatography to a stage showing a single major protein band of 70,000 daltons in a gel electrophoresis. The enzyme was capable of transcribing heteropolymeric RNA into DNA complements as demonstrated by specific back hybridization to template RNA. The leukemic spleen would appear to represent an important source of this enzyme, as well as other potentially important leukemia-specific reagents.

摘要

已从一名慢性淋巴细胞白血病患者的脾脏中制备出密度为1.16克/毫升、包裹着70S RNA和一种RNA指导的DNA聚合酶(逆转录酶)的颗粒。这些颗粒已被转化为密度为1.26克/毫升且含有酶-RNA复合物的核心,这与已知的禽源和鼠源RNA肿瘤病毒完全类似。通过柱色谱法从核心中纯化逆转录酶,直至在凝胶电泳中显示出一条70,000道尔顿的单一主要蛋白带。如通过与模板RNA的特异性反向杂交所证明的,该酶能够将杂聚RNA转录成DNA互补物。白血病脾脏似乎是这种酶以及其他潜在重要的白血病特异性试剂的重要来源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06e0/433153/7b1ea2844ed9/pnas00053-0353-a.jpg

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