Genetic Engineering - People Server at UNCW

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 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


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