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Nonisotopic assays for reporter gene activity
1University of Texas, Galveston, Texas, USA.
Current Protocols in Molecular Biology
|February 12, 2008
Summary
This study presents two sensitive chemiluminescent assays for measuring reporter gene activity using firefly luciferase and beta-galactosidase. These methods offer a broad linear range and can be performed with standard laboratory equipment.
Area of Science:
- Molecular Biology
- Biochemistry
Background:
- Reporter gene assays are crucial for studying gene expression.
- Nonisotopic detection methods offer advantages in sensitivity and safety.
Purpose of the Study:
- To describe two nonisotopic chemiluminescent assays for quantifying reporter gene activity.
- To evaluate the performance of firefly luciferase and beta-galactosidase reporter systems.
Main Methods:
- Cells were transfected with firefly luciferase or beta-galactosidase expression plasmids.
- Chemiluminescent detection of reporter gene activity was performed.
- Measurements were taken using a luminometer or scintillation counter.
Main Results:
- Both assays demonstrated high sensitivity.
- A broad linear range was observed for both reporter systems.
- Consistent results were obtained using either a luminometer or scintillation counter.
Conclusions:
- Chemiluminescent assays provide a sensitive and versatile method for measuring luciferase and beta-galactosidase activity.
- These nonisotopic systems are suitable for various gene expression studies.
