【摘要】 单一载体,即可实现多种实验目的的Halotag多功能标记系统在生物学中有着广泛的应用。本文首先介绍了Halotag多功能标记系统的组成、反应机制和改造过程,并简要概括了Halotag配基的类型。根据Halotag配基功能的差异,对其应用分为三个方面进行介绍:(1)不同的Halotag荧光配基可以实现在亚细胞水平的时空特异成像、单分子水平上的超分辨率显微成像以及在组织水平上对小动物进行活体成像。(2)Halotag固相配基可以用于蛋白纯化、蛋白-蛋白以及蛋白-DNA相互作用检测等,具有比其它蛋白标签更加优异的效果。(3)借助疏水性的Halotag配基能够诱导目的蛋白的降解。应用该技术,不仅能提高工作效率,还有助于对多个实验结果进行关联分析,在活细胞尤其哺乳动物细胞中有着极好的应用前景。更多还原

【Abstract】 Halotag multifunctional reporter system increasingly becomes one of the most popular labelling systems due to their properties of"one construct,and endless analysis". Firstly,we review the component,reaction mechanism and modification process of Halotag reporter system. According to the diversity of Halotag ligand,its applications are introduced as follows:( 1) we can use the Halotag fluorescent ligand to monitor the cellular process at subcellular level,single molecule level and tissue level.( 2) Halolink resin or Halochip can be used to purify protein or to capture cross-linked protein-DNA / protein-protein complexes.( 3) Introduction of hydrophobic moiety to Halotag fusion proteins can destine these proteins for effective proteasomal degradation. By means of this system, we will not only enhance our efficient,but facilitate data correlation across a series of related experiments as well. It is clear that Halotag multifunctional reporter system faces a bright future in living cells especially in mammalian cells.更多还原