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Genetic Engineering

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Genetic Engineering. Biotechnology Molecular Cloning Recombinant DNA. HISTORY. Stanley Cohen Competent Cells Plasmids. Herb Boyer EcoR1. Stanley Falkow. Asilomar Conference 1975. Voluntary Band of Some Recombinant Research NIH-RAC formed as a result. Isolate Gene. - PowerPoint PPT Presentation

Text of Genetic Engineering

  • Genetic Engineering

    BiotechnologyMolecular CloningRecombinant DNA

  • HISTORYHerb BoyerEcoR1Stanley CohenCompetent CellsPlasmidsStanley Falkow

  • Asilomar Conference 1975Voluntary Band of Some Recombinant ResearchNIH-RAC formed as a result

  • Isolate GeneFragmented Total DNA

    Use mRNA and Make DNA with Reverse Transcriptase

  • Splice into Cloning VehiclePlasmidsVirusesUse Restriction Endonuclease to create Sticking EndsRejoin Molecule withLigase

  • Inserting into Host

    TransformationDetection of Successful CloneDNA LibraryShotgun Cloning

  • Cloning VectorPlasmid Cloning VectorsSingle Cleavage SiteHigh copy NumbersInsertion Inactivation

  • Virus Vectors

    Lambda Phage VectorStable PackagingCosmids

  • Other VectorsExpression VectorsSecretion VectorsShuttle VectorsBacteriophage M13DNA SequencingYeast Artificial ChromosomeHuman Genome Project

  • Hosts

    Cloning Vector HostsE. coliEndotoxinsYeast

  • Detection of Clones

    AntibodiesNucleic Acid ProbesSynthetic DNA

  • Production of Recombinant ProteinTranscription of Cloned GenePromoter

    Translation of Cloned GeneRibosome Binding Site

  • Polymerase Chain Reaction

    Kary MullinsPCRDNA AmplificationTAQ EnzymePrimer

  • PCR

  • Site Directed MutagenesisKnockout Mutation

  • Applications of Genetic Engineering

    Human Insulin

    Interferon

    Hepatitis Vaccine

  • VaccinesSubunit VaccinesHepatitis VaccineRecombinant VaccinesPolyvalent VaccinesDNA VaccinesEdible Vaccines

  • Other ProductsTissue Plasminogen ActivatorRenninTi PlasmidBacillus thuringiensis BTRoundup ResistantFavosavr Tomato

  • More PossibilitiesIce Minus BacteriaPCB Degrading BacteriaLuminescent Reporter GeneTransgenic AnimalHuman Gene Therapy

  • DNA Chips - MicroarraysActual DNA Chip

  • Genetic Eng. Word ListBiotechnologyMolecular CloningRecombinant DNAIsolate Gene mRNA, Reverse Transcriptase Splice into Cloning Vehicle - Plasmids, Viruses Restriction Endonucleases Sticky Ends LigaseInsert into HostDetect Successful Clone DNA LibraryShotgun CloningPlasmid Cloning VectorsSingle Cleavage Sites

  • Genetic Eng. Word List 2High Copy numbersInsertion InactivationLambda Phage VectorStable PackagingCosmids Other VectorsExpression VectorsSecretion Vectors Shuttle VectorsBacteriophage M 13DNA SequencingYeast Artificial Chromosomes Human Genome ProjectCloning Vector HostsE. coliYeastDetection of ClonesAntibody

  • Genetic Eng. Word List 3Nucleic Acid ProbesTranscription of Cloned GenePromoterTranslation of Cloned GeneRibosome Binding SiteSynthetic DNAProbes

    Polymerase chain Reaction PCRDNA AmplificationTAQ EnzymePrimersSite Directed MutagenesisKnockout Mutations

    Applications of Genetic EngineeringHuman InsulinInterferonHepatitis VaccineRecombinant VaccineSubunit Vaccines

  • Genetic Eng. Word List 4Polyvalent VaccinesDNA VaccinesTissue Plasminogen ActivatorRenninTi PlasmidBacillus thuringiensis Roundup ResistanceFavosavr TomatoIce Minus BacteriaPCB Degrading BacteriaLuminescent Reporter GeneTransgenic Animals

    Edible VaccinesHuman Gene TherapyDNA Chips Microarrays