Reporter
Genes
Used
as
Scorable
Markers
and
Their
Enzyme
Assays
Reporter gene for
|
Substrate * and assay | Identification |
1. Chloramphenicol acetyl- transferase (CAT) |
(14C) chloramphenicol + acetyl CoA; TLC separation |
Detection of acetyl chloremphenicol by autoradiography |
| 2. Neomycin phosphotrans- ferase (NPT II) | Kanamycin + e2p) A TP (in situ assay) |
Radioactivity detection (also dot blots) |
| 3. β-glucuronidase (GUS) | Glucuronides (PNPG, x-GLUC, NAG, REG) |
Fluorescence detection: colourimetric, fluorometric and histochemical |
| 4. β-galactosidase (lac Z) | β -galactoside (X-gal)
|
Colour of the colony |
5. Luciferase |
(a) Decanal and FMNHz (b) ATP +02 +Iuciferin |
Bioluminiscence (exposure of x-rays film) |
6. Octopine synthase (OCS)
|
Ariginine + pyruvate + NADH |
Electrophoresis |
| 7. Nopaline synthase (NOS) | Arginine + ketoglutaric acid + NADH | Electrophoresis |