"I am that merry wanderer of the night"
- A Midsummer Nights Dream
Luciferase is the enzyme which is used by fire flies to glow. It uses the substrate luciferin for this oxidative process. The reaction is as seen below:
|University of Wisconsin, Dana Nystrom, http://people.uwec.edu/nystrodl/chem.htm|
So what are the advantages to using Luciferase versus traditional practices?
A close up of the main molecule
The opening shows how the substrate enters the enzyme. This is the opening to the active site.
What are things being done with Luciferase?
Renilla Luciferase (R-Luc) produces light upon mechanical stimulation. In this study R-Luc is fused to an antibody for cancer cells. The antibody used is Carinoembryonic antigen (CEA) was because it is highly expressed in colon cancer and is a surface marker. Fusion protein was tested for for binding to cancer cells through flow cytometry. The fusion protein was tested for expression and stability using bacterial periplasmic secretions and stability was improved by mutagenesis.
The protein was injected into a mouse under anesthesia, through the tail and PET and Optical imaging was used to capture the light emitted and the target tumor. The result was is as shown in the below picture:
Phase-Specific Circadian Clock Regulatory Elements in Arabidopsis
In this experiment, arabidopsis was transformed with Luciferase to study the Circadian biological clock. That is in plants, stoma opening and leaf movements are dependent on the biological clocks. The transformed arabidopsis were exposed to environmental conditions such as temperature and light/no light. Luciferase activity was then measured to help determine the activity of the plant. The experiment also included variations of the promoter region to determine which ones were important for circadian rhythms. They also were able to figure out which regions dealt with morning rhythms versus night rhythms. These rhythms were found to be most important on the transcriptional level.
With the ability to transform plants to glow like this, will Christmas Trees be next?