Scoreable Markers, Enzyme Assays, Choramphenicol Acetyltransferase, Kanamycin, Neomycin Phosphotrasferase, Luciferase, Octopine Synthase, Nopaline

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Gene Transfer Methods in Plants Gene Transfer Methods in Plants Introduction Target Cells for Transformation Theoretical Routes for Production of Transgenic Plants Vectors of Gene Transfer Vectors based on Ti and Ri Plasmids of Agrobacterium Structure and Function of Ti and Ri Plasmids Selectable Marker Genes in Vectors Used for Gene Transfer T-DNA Transfer Process Vectors based on Ti and Ri Plasmids Virus Vectors Transformation Techniques Using Agrobacterium Prerequisites for Production of Transgenic Plants Explants Used for Transformation Marker Genes for Selection and Scoring of Tranformed Cells/Calli or Shoots Reporter Genes Used as Scoreable Markers and their Enzyme Assays Neomycin Phosphotransferase Gene (nptII) Chloramphenicol Acetyl Transferase Gene (cat)

Home >> Plant Biotechnology and Genomics >> Gene Transfer Methods in Plants >>Reporter Genes used as Scoreable Markers, and their Enzyme Assays

Reporter Genes used as Scoreable 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
2. Neomycin phosphotransferase (NPTII)	Kanamycin + ³²P) ATP (in situ assay)	chloramphenicol by autoradiography
3. b -glucuronidase (GUS; uidAgene)	Glucuronides (PNPG; x-GLUC, NAG, REG)	Radioactivity detection (also dot blots)
4. b-galactosidase (lac Z)	b-galactoside (X-gal)	Fluorescence detection: colorimetric, fluorometric and histochemical
5. Luciferase	(a) Decanal and FMNH₂ (b) ATP + O₂+ luciferin	Colour of the colony
6. Octopine synthase (OCS)	Ariginine + pyruvate + NADH	Bioluminiscence (exposure of x-rays film)
7. Nopaline synthase (NOS)	Arginine + ketoglutaric acid + NADH	Electrophoresis
8. Green fluorescent protein (GFP)	No substrate is needed	Emission of green light when exposed to UV light

*PNPG = p-nitrophenyl glucuronide ; x-GLUC = 5-bromo, 4-chloro, 3-indolyl glucuronide; NAG = naphthol AS-BI glucuronide ; REG = Resorufin glucuronide.

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β-Glucuronidase (GUS)Gene (uidA) Luciferase Gene (lux) Green Fluorescent Protein Gene Phosphomannose Isomerase Gene (pmi) Agrobacterium-Mediated Transformation in Monocots Agroinfection and Gene Transfer Physical Delivery or DNA Mediated Gene Transfer (DMGT) Chemically Stimulated DNA Uptake by Protoplasts Microinjection and Macroinjection Microprojectiles for Gene Transfer Electroporation Method for Gene Transfer Liposome Mediated Gene Transfer Calcium Phosphate Precipitation Method Transformation Using Pollen or Pollen Tube Incubation of Dry Seeds,Embryos,Tissues or Cell in DNA Transformation by Ultrasonication Removal of Selectable Marker Gene from Transgenic Plants Need for Removal of Selectable Marker Gene Genetic Systems Used for Removal of Marker Gene Examples of North American Plant Transformation Facilities Plant Transformation Facilities (PTFs)