Chemically Stimulated DNA, Protoplasts, Liposomes, Protoplast System, Polythylene Glycol, Macromolecules, Endocytosis, Petunia, Necotiana

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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 >>Chemically Stimulated DNA Uptake by Protoplasts

Chemically Stimulated DNA Uptake by Protoplasts
Direct. DNA uptake by protoplasts can be stimulated by chemicals like polyethylene glycol (PEG). The technique is so efficient that virtually every protoplast system has proved transformable. PEG is also used to stimulate the uptake of liposomes and to improve the efficiency of e1ectroporation (see below for details). PEG at high concentration (15-25%) will precipitate ionic macromolecules such as DNA and stimulate their uptake by endocytosis without any gross damage to protoplasts. This is followed by cell wall formation and initiation of cell division. These cells can now be plated at low density on selection medium.

Initial studies using the above method were restricted to Petunia and Necotiana.However, other plant systems (rice, maize, etc.) were also successfully used later. In these methods, PEG was used in combination with pure Ti plasmid, or calcium phosphate precipitated Ti plasmid mixed with a carrier DNA. Transformation frequencies upto 1 in 100 have been achieved by this method. Nevertheless, there are serious problems in using this method for getting transgenic plants, mainly due to difficulties encountered in plant regeneration from protoplasts.

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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)