Genetic Engineering

• Biotechnology• Molecular Cloning• Recombinant DNA

HISTORY

Herb Boyer

EcoR1

Stanley Cohen

Competent Cells

Plasmids

Stanley Falkow

Asilomar Conference 1975

Voluntary Band of Some Recombinant Research

NIH-RAC formed as a result

Isolate Gene

• Fragmented Total DNA

• Use mRNA and Make DNA with Reverse Transcriptase

Splice into Cloning Vehicle

• Plasmids• Viruses• Use Restriction

Endonuclease to create –

• Sticking Ends• Rejoin Molecule with• Ligase

Inserting into Host

• Transformation• Detection of

Successful Clone• DNA Library• Shotgun Cloning

Cloning Vector

• Plasmid Cloning Vectors

• Single Cleavage Site

• High copy Numbers• Insertion Inactivation

Virus Vectors

• Lambda Phage Vector

• Stable Packaging• Cosmids

Other Vectors

• Expression Vectors• Secretion Vectors• Shuttle Vectors• Bacteriophage M13• DNA Sequencing• Yeast Artificial

Chromosome• Human Genome Project

Hosts

• Cloning Vector Hosts

• E. coli• Endotoxins• Yeast

Detection of Clones

• Antibodies• Nucleic Acid Probes• Synthetic DNA

Production of Recombinant Protein

• Transcription of Cloned Gene

• Promoter

• Translation of Cloned Gene

• Ribosome Binding Site

Polymerase Chain Reaction

• Kary Mullins• PCR• DNA Amplification• TAQ Enzyme• Primer

PCR

Site Directed Mutagenesis

Knockout Mutation

Applications of Genetic Engineering

• Human Insulin

• Interferon

• Hepatitis Vaccine

Vaccines

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

Other Products

• Tissue Plasminogen Activator

• Rennin• Ti Plasmid• Bacillus

thuringiensis – BT• Roundup Resistant• Favosavr Tomato

More Possibilities

• Ice Minus Bacteria• PCB Degrading

Bacteria• Luminescent Reporter

Gene• Transgenic Animal• Human Gene Therapy

DNA Chips - Microarrays

Actual DNA Chip

Genetic Eng. Word List

Biotechnology

Molecular Cloning

Recombinant DNA

Isolate Gene

mRNA,

Reverse Transcriptase

Splice into Cloning Vehicle -

Plasmids, Viruses

Restriction Endonucleases

Sticky Ends

Ligase

Insert into Host

Detect Successful Clone

DNA Library

Shotgun Cloning

Plasmid Cloning Vectors

Single Cleavage Sites

Genetic Eng. Word List 2

High Copy numbers

Insertion Inactivation

Lambda Phage Vector

Stable Packaging

Cosmids

Other Vectors

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

Genetic Eng. Word List 3Nucleic 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

Genetic Eng. Word List 4

Polyvalent Vaccines

DNA Vaccines

Tissue Plasminogen Activator

Rennin

Ti Plasmid

Bacillus thuringiensis

Roundup Resistance

Favosavr Tomato

Ice Minus Bacteria

PCB Degrading Bacteria

Luminescent Reporter Gene

Transgenic Animals

Edible Vaccines

Human Gene Therapy

DNA Chips – Microarrays