DNA is universal. Long history of manipulating genes Speed- horses Strength- oxen Larger fruit.

Genetic Engineering

DNA is universal

Selective Breeding Long history of

manipulating genes Speed- horses

Strength- oxen

Larger fruit

Science of Genetic Engineering

Moving genes from one organism to another.

Based upon genetics

Began in the 1970’s- manipulations of bacteria, viruses and plasmids

Processes of Genetic Engineering

1. Recombinant DNA2. DNA Fingerprinting3. Polymerase chain reaction4. Cloning

Chimera

First Chimera1973- Cohen & Boyer

Isolated the gene – for rRNA from an

African clawed toad

Inserted it into a bacterium

Tools of Genetic Engineering

Restriction enzymes

Ligase

Vectors

Gel electrophoresis

PCR

Restriction Enzymes- endonucleases

Discovered in 1968- origin Bacteria

Chemical scissors

Used as a defense mechanisms against foreign DNA.

100’s have been

identified

Bind to a specific sequence (4-6) nucleotides.

They cut the DNA leaving sticky ends

EcoRI

Named for the organism of origin.

Restriction Enzymes- blunt ends

Hemophilus aegypticus ( HaeIII)

LigaseAn enzyme that

binds nucleotides together

“Super glue” – genetic engineering

Vectors

Something that delivers a biological material from one organism to another.

Plasmids

Bacteria

Extra DNA – separate from the main genome (nucloid)

Ability to replicate

1,000 – 20,000 base pairs

VirusesViruses have evolved

mechanisms to transport their genomes into a host cell.

Transduction- delivery of a gene by a virus.

Pathogenic viruses are modified to minimize risk.

Gene guns and metal bullets

Delivery system designed for injecting cells with genetic information.

“biolistic particle delivery system”

Payload is an elemental particle coated with a plasmid.

Recombinant DNAThe process of

creating a chimera.

“Splicing a gene/genes from one organism into the genome of a different organism”.

Paper Recombinant DNA

Genetically Modified OrganismGMO

Organism that has acquired 1 or more genes by artificial means

New be from another species

Transgenic- an organism contains a gene from another organism.

BacteriaOften the organism

of choice:1. Grow quickly2. Low costs3. Easily engineered

to secrete into a growing medium

H. Insulin- Humulin 1982

H growth hormone Epidermal growth

hormone Interleukin2 –

cancer Taxol – cancer

Yeast

Eukaryotes Single celled Contain plasmids

Yeast are:

Easy to grow Take up foreign

DNA Better at

synthesizing and secreting eukaryotic proteinsHepatitis B

vaccineInterferon- cancer, viruses

“Pharm” animalsMAMMALS

Many of the proteins that are secreted by humans are glycoproteins.

Only mammals can correctly add the sugar to these proteins.

When a gene of interest is inserted into a specific mammary cell= it will be secreted into the milk.

Milk medicine

1. Erythropoitin- anemia

2. Factor VIII- hemophilia

Genetically modified organisms

GoatsGoats contain gene

that codes for spider silk.

Extracted from milk.

Bullet proof vests Artificial tendons clothing

Tumor resistant mouse Contains a gene

that codes for a protein called par-4

Kills cancer cells not normal cells.

Emerald Sea Slug Believed to be the first

known, naturally occurring transgenic animal.

Feeds on algae- capable of using the algae to carry out photosynthesis.

Found a gene in the slug with the same genetic sequence as algae.

l

Glofish 1st commercial

transgenic animal Contain a gene

from jellyfish that allow cells to fluoresce

Aedes mosquito Genetically

engineered to be sterile

Mate with females Population of

mosquitos reduced.

Less disease like dengue fever that has no vaccine or cure.

Mighty mouse Genetically altered

to over-produce the enzyme PEPCK-C involved in glucose generation.

Runs 25x longer

Affects on aging

Transgenic chickens Able to express

foreign products in their eggs.

Goal for chickens to produce eggs with life saving drugs.

GM and AgricultureHow to feed a

growing population?

Use a soil bacterium :

Agrobacteria tumifaciens

Ti plasmid

ApplicationsMost Soybeans and

cotton crops are GM.

Pesticide resistant Glycophosphate

resistant “round-up”

Pest resistant

To improve nutritional value of crops.

Golden rice- contain daffadil gene ( beta-carotene)-

reduce blindness

Risks of GM food

Animals: What if a cancer cell

was transferred to a virus or bacterium.

Crops: Allergies Gene drift to nearby

crops. Hazardous to human

health

Government Regulations

Strict laboratory procedures

Genetically altered microbes must be

disabled. Dangerous

experiments have been banned

BioremediationBacteria- clean up

wastes.1. Take up mercury-

pollution2. Extract pesticide

vapors from the air

3. Take up petroleum from spills

EnviropigEngineered to

produce and enzyme in its saliva which breaks down phosphorus in their feces.

Reduces pollution Marine pollution-

runoff Greenhouse gases

Gel electrophoresis Acts like a sieve to

separate out DNA fragments by size.

Separates linear pieces of DNA into bands.

Length : distance

RFLP’s DNA from different people- cut with

the same restriction enzyme =

RFLP’s- restriction fragment length polymorphisms.

Electrophoresis- generates a fingerprint that shows relationship between organisms.

Application of DNA Fingerprints Determine paternity- T.

Jefferson – Sally Hemings

Forensics- solving cold cases

Identifying endangered species trafficing

Identify genes that cause genetic disorders

billion copies of a DNA – few hours

Simple process: DNA DNA polymerase Nucleotide

monomers Heating and

cooling

Ancient DNA – wholly mammoth

Forensics- blood, semen, tissue

Prenatal diagnosis of genetic disorders

DNA from viral genes from infected cells

PCR- Polymerase Chain Reaction