Unbiased tracking of the progression of mRNA and protein synthesis in bulk and in liposome-confined reactions

Pauline van Nies, Zohreh Nourian, Maurits Kok, Roeland van Wijk, Jonne Moeskops, Ilja Westerlaken, Jos M. Poolman, Rienk Eelkema, Jan H. van Esch, Yutetsu Kuruma, Takuya Ueda, Christophe Danelon*

*この研究の対応する著者

研究成果: Article査読

30 被引用数 (Scopus)

抄録

The compartmentalization of a cell-free gene expression system inside a self-assembled lipid vesicle is envisioned as the simplest chassis for the construction of a minimal cell. Although crucial for its realization, quantitative understanding of the dynamics of gene expression in bulk and liposome-confined reactions is scarce. Here, we used two orthogonal fluorescence labeling tools to report the amounts of mRNA and protein produced in a reconstituted biosynthesis system, simultaneously and in real-time. The Spinach RNA aptamer and its fluorogenic probe were used for mRNA detection. Applying this dual-reporter assay to the analysis of transcript and protein production inside lipid vesicles revealed that their levels are uncorrelated, most probably a consequence of the low copy-number of some components in liposome-confined reactions. We believe that the stochastic nature of gene expression should be appreciated as a design principle for the assembly of a minimal cell.

本文言語English
ページ(範囲)1963-1966
ページ数4
ジャーナルChemBioChem
14
15
DOI
出版ステータスPublished - 2013 10
外部発表はい

ASJC Scopus subject areas

  • 生化学
  • 分子医療
  • 分子生物学
  • 有機化学

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