Monitoring Promoter Activity by Flow Cytometry.

ABSTRACT

Genetic reporters have become invaluable tools for indirectly monitoring promoter activities. The quantitative measurement of promoter activities using reporter gene systems is fundamental for pharmaceutical, biomedical, and molecular biology research. Genetic reporters are used not only for measuring promoter activities but also for understanding the mechanisms controlling gene transcription and in the identification, and characterization of cis-acting regulatory elements. Fluorescent reporter proteins including enhanced green fluorescent protein (EGFP ) are reliable for monitoring quantitative underlying differences in promoter activities. The emitted fluorescence intensity of the expressed reporter is measured at the single-cell level by flow cytometry and represents a readout for the promoter activities. In this chapter, the protocol for measurement and analyzing of transfected cells expressing the reporter gene EGFP is thoroughly described and fully illustrated.