生物正交化学
标签
化学
蛋白质标签
点击化学
转移酶
生物化学
叠氮化物
体内
基质(水族馆)
体外
组合化学
肉豆蔻酰化
融合蛋白
重组DNA
酶
有机化学
膜
生物
生态学
生物技术
基因
作者
William P. Heal,Sasala R. Wickramasinghe,Robin J. Leatherbarrow,Edward W. Tate
摘要
N-Myristoyl transferase-mediated labelling using a substrate modified with an azide or alkyne tag is described as an efficient and site-selective method for the introduction of a bioorthogonal tag at the N-terminus of a recombinant protein. The procedure may be performed in vitro, or in a single over-expression/tagging step in vivo in bacteria; tagged proteins may then be captured using Staudinger-Bertozzi or 'click' chemistry protocols to introduce a secondary label for downstream analysis. The straightforward synthesis of the chemical and molecular biological tools described should enable their use in a wide range of N-terminal labelling applications.
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