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Genetic Engineering

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.

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Genetic Engineering

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  1. Genetic Engineering • Biotechnology • Molecular Cloning • Recombinant DNA

  2. HISTORY Stanley Cohen Competent Cells Plasmids Herb Boyer EcoR1 Stanley Falkow

  3. Asilomar Conference 1975 Voluntary Band of Some Recombinant Research NIH-RAC formed as a result

  4. Isolate Gene • Fragmented Total DNA • Use mRNA and Make DNA with Reverse Transcriptase

  5. Splice into Cloning Vehicle • Plasmids • Viruses • Use Restriction Endonuclease to create – • Sticking Ends • Rejoin Molecule with • Ligase

  6. Inserting into Host • Transformation • Detection of Successful Clone • DNA Library • Shotgun Cloning

  7. Cloning Vector • Plasmid Cloning Vectors • Single Cleavage Site • High copy Numbers • Insertion Inactivation

  8. Virus Vectors • Lambda Phage Vector • Stable Packaging • Cosmids

  9. Other Vectors • Expression Vectors • Secretion Vectors • Shuttle Vectors • Bacteriophage M13 • DNA Sequencing • Yeast Artificial Chromosome • Human Genome Project

  10. Hosts • Cloning Vector Hosts • E. coli • Endotoxins • Yeast

  11. Detection of Clones • Antibodies • Nucleic Acid Probes • Synthetic DNA

  12. Production of Recombinant Protein • Transcription of Cloned Gene • Promoter • Translation of Cloned Gene • Ribosome Binding Site

  13. Polymerase Chain Reaction • Kary Mullins • PCR • DNA Amplification • TAQ Enzyme • Primer

  14. PCR

  15. Site Directed Mutagenesis Knockout Mutation

  16. Applications of Genetic Engineering • Human Insulin • Interferon • Hepatitis Vaccine

  17. Vaccines • Subunit Vaccines • Hepatitis Vaccine • Recombinant Vaccines • Polyvalent Vaccines • DNA Vaccines • Edible Vaccines

  18. Other Products • Tissue Plasminogen Activator • Rennin • Ti Plasmid • Bacillus thuringiensis – BT • Roundup Resistant • Favosavr Tomato

  19. More Possibilities • Ice Minus Bacteria • PCB Degrading Bacteria • Luminescent Reporter Gene • Transgenic Animal • Human Gene Therapy

  20. DNA Chips - Microarrays Actual DNA Chip

  21. Genetic Eng. Word List Splice into Cloning Vehicle - Plasmids, Viruses Restriction Endonucleases Sticky Ends Ligase Insert into Host Biotechnology Molecular Cloning Recombinant DNA Isolate Gene mRNA, Reverse Transcriptase Detect Successful Clone DNA Library Shotgun Cloning Plasmid Cloning Vectors Single Cleavage Sites

  22. Genetic Eng. Word List 2 Other Vectors Expression Vectors Secretion VectorsShuttle Vectors Bacteriophage M 13 DNA Sequencing Yeast Artificial Chromosomes Human Genome Project Cloning Vector Hosts E. coli Yeast Detection of Clones Antibody High Copy numbers Insertion Inactivation Lambda Phage Vector Stable Packaging Cosmids

  23. Genetic Eng. Word List 3 Nucleic Acid Probes Transcription of Cloned Gene Promoter Translation of Cloned Gene Ribosome Binding Site Synthetic DNA Probes Polymerase chain Reaction – PCR DNA Amplification TAQ Enzyme Primers Site Directed Mutagenesis Knockout Mutations Applications of Genetic Engineering Human Insulin Interferon Hepatitis Vaccine Recombinant Vaccine Subunit Vaccines

  24. Genetic Eng. Word List 4 Polyvalent Vaccines DNA Vaccines Tissue Plasminogen Activator Rennin Ti Plasmid Bacillus thuringiensis Edible Vaccines Human Gene Therapy DNA Chips – Microarrays Roundup Resistance Favosavr Tomato Ice Minus Bacteria PCB Degrading Bacteria Luminescent Reporter Gene Transgenic Animals

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