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Genetic Engineering Biotechnology Molecular Cloning Recombinant DNA.

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Presentation on theme: "Genetic Engineering Biotechnology Molecular Cloning Recombinant DNA."— Presentation transcript:

1 Genetic Engineering Biotechnology Molecular Cloning Recombinant DNA

2 HISTORY Stanley Falkow Stanley Cohen Herb Boyer Competent Cells EcoR1
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 - Biotechnology
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 Human Genome Project
Expression Vectors Secretion Vectors Shuttle 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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