Enzyme-generated intracellular fluorescence for single-cell reporter gene analysis utilizing Escherichia coli β-glucuronidase
✍ Scribed by Matthew Lorincz; Mario Roederer; Zhenjun Diwu; Leonard A. Herzenberg; Garry P. Nolan
- Book ID
- 102651527
- Publisher
- John Wiley and Sons
- Year
- 1996
- Tongue
- English
- Weight
- 907 KB
- Volume
- 24
- Category
- Article
- ISSN
- 0196-4763
No coin nor oath required. For personal study only.
✦ Synopsis
We report the development of a new fluorescence-activated cell sorter (FACS)-based reporter gene system utilizing the enzymatic activity of the E. coZi P-glucuronidase @us) gene. When loaded with the Gus substrate fluorescein-di-P-D-glucuronide (FDGlcu), individual mamma&m cells expressing and translatinggus mRNA liberate s&cient levels of intracellular fluorescein for quantitative analysis by flow cytometry. This assay can be used to FACS sort viable cells based on Gus enzymatic activity, and the efficacy of the assay can be 0 1996 Wiley-Llss, I~c.