SpringerProtocols: Abstract: Use of Liposome-Mediated DNA Transfection to Determine Promoter Activity in Smooth Muscle Cells

Use of Liposome-Mediated DNA Transfection to Determine Promoter Activity in Smooth Muscle Cells

By: Rosalind P. Fabunmi²

Abstract

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The transfer and expression of DNA plasmids containing promoter fragments of heterologous genes linked to reporter cDNAs in mammalian cells has become an invaluable technique for studying the regulation of gene expression. Several reporter genes such as luciferase, β-galactosidase, chloramphenicol acetyl transferase, and green flourescent protein are ideal to study promoter activities as their gene products are not endogenous to smooth muscle cells (SMC) and their expression can be readily detected using convenient assays (1). Among these genes, a popular choice is the firefly luciferase, as its expression can be easily detected in cells using a highly sensitive chemiluminescent assay (2). The firefly luciferase catalyses a rapid, ATP-dependent oxidation of the substrate, luciferin, which then emits light. Reactions catalyzed by firefly luciferase are:

Affiliation(s): (2) Department of Physiology, University of Texas Southwestern Medical Center, Dallas, TX

Book Title: Vascular Disease: Molecular Biology and Gene Therapy Protocols

Series: Methods in Molecular Medicine | Volume: 30 | Pub. Date: Aug-01-1999 | Page Range: 143-150 | DOI: 10.1385/1-59259-247-3:143

Subject: Molecular Medicine

References

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