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天然和重组双链RNA特异性腺苷脱氨酶的纯化。

Purification of native and recombinant double-stranded RNA-specific adenosine deaminases.

作者信息

O'Connell M A, Gerber A, Keegan L P

机构信息

MRC Human Genetics Unit, Western General Hospital, Edinburgh, United Kingdom. hgu.mrc.ac.uk

出版信息

Methods. 1998 May;15(1):51-62. doi: 10.1006/meth.1998.0605.

Abstract

ADAR1 and ADAR2 are members of a family of enzymes that catalyze the conversion of adenosine to inosine in double-stranded RNA. Unlike the other types of RNA editing that involve multiprotein editing complexes, the site-specific deamination of an adenosine to inosine is catalyzed by single enzymes. ADAR1 and ADAR2 have been purified and the genes cloned from various sources. Each gene encodes multiple splice variants. As it is crucial to have an adequate supply of pure protein to investigate this type of RNA editing, we describe in this article methods for both the purification and the overexpression of either full-length or partial ADAR1 and ADAR2 isoforms.

摘要

ADAR1和ADAR2是一类酶家族的成员,这类酶催化双链RNA中的腺苷向肌苷的转化。与其他涉及多蛋白编辑复合体的RNA编辑类型不同,腺苷向肌苷的位点特异性脱氨基作用由单一酶催化。ADAR1和ADAR2已被纯化,其基因也已从各种来源克隆出来。每个基因编码多种剪接变体。由于拥有足够的纯蛋白供应对于研究此类RNA编辑至关重要,我们在本文中描述了全长或部分ADAR1和ADAR2亚型的纯化及过表达方法。

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