An in vivo translation-reporter system for the study of protein synthesis in zebrafish embryos.

ABSTRACT: Control of gene expression at the translation level is increasingly regarded as a key feature in many biological processes. Simple, inexpensive and reliable procedures to visualize sites of protein production are required to allow observation of the spatiotemporal patterns of mRNA translation at subcellular resolution. We present a method, named SPoT (for Subcellular Patterns of Translation), developed upon the original TimeStamp technique ( Lin et al., 2008), consisting in the expression of a fluorescent protein fused to a tagged, self-cleavable protease domain. The addition of a cell-permeable protease inhibitor instantly stabilizes newly produced tagged protein allowing us to distinguish recently synthesized proteins from pre-existing ones. After a brief protease inhibitor treatment, th

SUBMITTER: Garcez Palha I 

PROVIDER: S-EPMC6310896 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature