Analysis 2009 - Additional Transformation Topics...

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Additional Transformation Topics
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Chloroplast Transformation
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For “containing” transgenes (plastids not transferred through pollen?) Chloroplast transformation involves homologous recombination Leaf discs are bombarded with plasmid constructs containing a selectable antibiotic resistance marker physically linked to the gene of interest, flanked by DNA for inserting into the correct site of the chloroplast genome
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The antibiotic resistance marker most frequently used is the aadA gene encoding resistance for spectinomycin and streptomycin, driven by the promoter of the chloroplast encoded 16S rRNA gene. This transformation procedure applied to tobacco, Arabidopsis or oil seed rape, generates plants in which all the chloroplast genomes are uniformly transformed (a condition referred to as homoplasmic), despite the fact that tobacco leaf cells may contain 100 chloroplasts, each containing 100 copies of the chloroplast genome.
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Another advantage of chloroplast transformation
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