What is the DNA sequence of the luciferase gene?
What is the DNA sequence of the luciferase gene?
The luciferase gene was composed of seven exons, coding for 547 amino acids, separated by six introns and spanning 1976 bp of genomic DNA (Fig. 1 and Figure 2). The entire 3086 bp sequence, including primer sequence, was deposited in GenBank, accession # DQ072141.
How many amino acids does luciferase have?
Luciferase is a single-chain polymer comprised of 548 amino acid residues (RCSB Protein Data Bank, 2006).
What is the gene code for luciferin?
The h3h gene showed sequence similarity with 3-hydroxybenzoate 6-monooxygenases, enzymes that catalyze oxidation of 3-hydroxybenzoate using NADH and molecular oxygen. This reaction is identical to that which converts hispidin into luciferin (Fig.
How does luciferase assay work?
The luciferase assay is useful to study whether a protein of interest regulates a particular gene at the transcription level. When this protein activates transcription, the cell will produce luciferase enzyme. After the addition of a lysis buffer and a substrate, a luminometer quantifies the luciferase activity.
Where is luciferin from?
“Luciferin” got its name from the Latin word “lucifer” (meaning “light-bearing”), which is also a source of the word that is sometimes used as a name of the devil.
Is luciferase dimeric or monomeric?
(≥ 3 mg/ml) luciferase is predominantly dimeric (luciferase is approximately 50% dimeric at 3 mg/ml). At typical working concentrations (0.01 to 1 ng/ml) luciferase is primarily monomeric. The isoelectric point (pI) has been determined to be 6.7 and the pH optimum is 7.8.
Is luciferase a dimeric protein?
Firefly luciferase is extremely hydrophobic; the monomer is apparently active, but the enzyme exists as a dimer in most solutions [6].
How do you perform luciferase assay?
Luciferase Reporter Assay To perform the luciferase assay, 100 µ l of each plate lysate is added to 360 µ l of luciferase assay buffer (Table I) in a luminometer vial. The reactions are started by the injection of 200 µ l of the 0.2 mM luciferin solution (Table I), and light output is measured for 20 sec at 25°C.
