Virus Sensing and Signaling Unit, Department of Virology, Institut Pasteur, UMR3569 CNRS, Paris, France.
Université de Paris, Paris, France.
Methods Mol Biol. 2022;2503:195-204. doi: 10.1007/978-1-0716-2333-6_15.
Bioorthogonal labelling of living cells enables the incorporation of small, chemically inert units (alkynes or azides) into nascent chains of biomolecules allowing the tracking of DNA synthesis, transcription, and translation in a temporal-spatial manner without compromising their integrity. This chemical labelling method can be used to replace traditional radiolabelled nucleosides, ribonucleosides, or amino acids with the added benefit of enabling visualization using confocal or super-resolution microscopy. Here, we outline our recently published methods for labelling nascent DNA and polypeptides in cells infected with African swine fever virus.
活细胞的生物正交标记能够将小的、化学惰性的单元（炔烃或叠氮化物）掺入生物分子的新生链中，从而在不破坏其完整性的情况下，以时空方式跟踪 DNA 合成、转录和翻译。这种化学标记方法可以替代传统的放射性标记核苷、核糖核苷或氨基酸，并且具有使用共聚焦或超分辨率显微镜进行可视化的额外优势。在这里，我们概述了我们最近发表的用于标记感染非洲猪瘟病毒的细胞中新生 DNA 和多肽的方法。
