School of Life Science, Beijing Institute of Technology , Beijing 100081, China.
Anal Chem. 2017 Nov 7;89(21):11620-11627. doi: 10.1021/acs.analchem.7b03043. Epub 2017 Oct 16.
Though techniques in bioorthogonal chemistry and metabolic incorporation have been developed over the past decade, it remains difficult to integrate different bioorthogonal reactions or metabolic incorporations into one system. In this report, the protein and DNA metabolic incorporations were combined with two bioorthogonal reactions in one cell to develop a facile and universal method for virus dual labeling. Azide and vinyl groups were introduced into the proteins or genomes of viruses, respectively, through the intrinsic biosynthesis of biomolecules, which were subsequently fluorescently labeled via copper-free click chemistry or alkene-tetrazine ligation reactions during natural propagation process in host cells. Both the envelope viruses and the capsid viruses could be labeled, and the dual labeling efficiency was more than 80%. The labeled progeny virions were structurally intact and fully infectious, and their fluorescence was strong enough to track single virions.
尽管过去十年中已经开发出了生物正交化学和代谢掺入技术，但将不同的生物正交反应或代谢掺入整合到一个系统中仍然很困难。在本报告中，蛋白质和 DNA 的代谢掺入与两种生物正交反应在一个细胞中结合，开发了一种简便通用的病毒双重标记方法。通过生物分子的内在生物合成，分别将叠氮化物和乙烯基基团引入病毒的蛋白质或基因组中，然后在宿主细胞中的自然繁殖过程中通过无铜点击化学或烯炔-四嗪连接反应进行荧光标记。包膜病毒和衣壳病毒都可以被标记，双重标记效率超过 80%。标记的子代病毒粒子结构完整且具有完全感染力，其荧光强度足以跟踪单个病毒粒子。
