Vectors Gene Transfer, Marker Genes, Eukaryotic Promoter, Bacterial Antibiotic Resistance Gene, Origin Replication, Plant Genetic Engineering

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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)

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Vectors of Gene Transfer
General features
The common features of vectors used for transformation of eukaryotic cells are shown in. Most vectors carry marker 'genes, which allow recognition of transformed cells (other cells die due to the action of an antibiotic or herbicide) and are described as selectable markers.

(a) Generalised Vector for the Transformation of Eukaryotic Cells, (b) Details of the Selectable marker Genes (Redrawn from D. Grierson, 'Plant Genetic Engineering',1991)

(a) Generalised Vector for the Transformation of Eukaryotic Cells, (b) Details of the Selectable Marker Genes(Redrawn from D.Grierson, 'Plant Genetic Engineering',1991)

Among these marker genes, the most common selectable marker gene is nptlI,providing kanamycin resistance.

Other common features of a suitable transformation vector include the following: (i) multiple unique restriction sites (a synthetic polylinker); (ii) bacterial origins of replication (e.g. CoIEl). The vectors having these properties may not neccessarily have features, which facilitate their transfer to plant cells or integration into the plant nuclear genome. Therefore, Agrobacterium Ti plasmid is preferred over all other vectors, because of wide host range of this bacterial system and the
capacity to transfer genes due to the presence of T-DNA border sequences.

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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)