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Analysis of Transgenic Plants

Analysis of Transgenic Plants. Requirements. Sequence of gene of interest Gene construct. Gene construct. GMO detection system. Morphology Physiology Yield characters Confirmation selectable marker or gene of interest Gene expression Gene sequence. Selection based on

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Analysis of Transgenic Plants

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  1. Analysis of Transgenic Plants

  2. Requirements • Sequence of gene of interest • Gene construct

  3. Gene construct

  4. GMO detection system • Morphology • Physiology • Yield characters • Confirmation selectable marker or gene of interest • Gene expression • Gene sequence

  5. Selection based on • Selectable Marker Gene (Morphology)

  6. Protein Based Testing Methods • Transgenes encode for novel proteins • Immunoassay techniques using antibodies • Ideal for qualitative and quantitative needs • Detects specific proteins in complex matrices • Analyte must be known • Interference from non-specific interactions of proteins, surfactants (saponins), phenolics, fatty acids and phosphatases • Polyclonal antibodies • Sensitive but less specific recognition • Monoclonal antibodies • Highly specific, slightly less sensitive recognition

  7. 35S promoter EPSPS CP4 NOS terminator CaMV Agrobacterium tumefaciens • Typically included in rDNA: promoter sequence, structural gene and a stop sequence DNA-Based Methods • Rely on 2 complementary DNA strands that hybridize in a sequence-specific manner • rDNA in the crop consists of several unique elements

  8. Southern Analysis

  9. Principles of PCR • Allows millifold amplification of target DNA • DNA polymerase makes exact copy of template • Synthetic oligonucleotide primers • Frame desired target sequence • Complimentary to DNA • Allows Taq-polymerase enzyme to generate complimentary sequence between primer sets • Sequence # grows exponentially • In theory…

  10. Progeny Testing

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