Transgenic organisms or

Genetically ModifiedOrganisms (GMOs)

Review of recombinant DNA technology

Growth Hormone

Reading for this and next lecture: Chapter 19

GAATTC

CTTAAG

G

CTTAA

AATTC

G

EcoRI

Restriction enzymes cut DNA a specific sites.

e.g., EcoRI recognizes GAATCC.

GAATTC

CTTAAG

GAATTC

CTTAAG

G

CTTAA

AATTC

G

G

CTTAA

AATTC

G

EcoRI cut

Mix and ligate

G

CTTAA

AATTC

G

G

CTTAA

AATTC

G

Recombinant

molecules

GAATTC

CTTAAG

GAATTC

CTTAAG

Parental

molecules

Cut two DNAs with EcoRI, mix and ligate.

Voila, recombinant DNA!

Sealing of the broken

phosphodiester bond

is called ligation and

performed with an

enzyme called DNA

ligase.

Small protein (191 amino acids) expressed fromanterior lobe of pituitary

Essential for normal development

Promotes growth, especially in children

Growth Hormone (GH)

Mutations that eliminate the function GH1 generesult in severe dwarfism

Too much GH in childhood results in gigantismRobert Pershing Wadlow

Too much GH in adults resultsin acromegaly

GH

Activated GH receptor

Inactive GH receptor

Binding of GH to cell-surface receptor leads to

transcriptional regulation of specific genes.

Transcription factor

GH target gene

Injected growth hormone can be used to treat:

*growth hormone deficiency (GHD) in pediatric patients

*growth failure associated with chronic renal insufficiency(CRI) prior to kidney transplantation

*short stature associated with Turner syndrome

*growth hormone deficiency (GHD) in adults

In the 1960s and 70s, children were treated withgrowth hormone isolated from human cadavers.(80 cadavers for 1 year of treatment)

In 1985, the FDA banned this practice becauseseveral patients died from a fatal brain disorder

known as Creutzfeldt-Jacob Disease (CJD). CJDwas caused by contaminating prions in thegrowth hormone extract.

In 1985, Genentech introduced Protropin, arecombinant GH made in bacteria.

Some GH history How was human growth hormone(HGH) produced in bacteria?

Make DNA copy of hGH mRNA. This copy isknown as a complementary DNA or cDNA.

Clone cDNA into plasmid.

Introduce recombinant plasmid in bacteria (E.coli).

Induce expression of gene and purify HGH.

BALCO Chief on Sports Doping Scandal

Victor Conte Tells '20/20' He Created Illegal Drug Regimens for Top AthletesDec. 3, 2004 — Victor Conte, a man at the center of an anti-doping scandal that has rocked the toptiers of the sports world, tells "20/20" he tailored illegal drug regimens for top athletes, includingOlympic track stars Marion Jones, Kelli White and Tim Montgomery.

.

.Conte, founder of the California-based Bay Area Laboratory Co-Operative, which is facing federalcharges for distributing steroids and other illegal performance-enhancing drugs to athletes, sayshe supplied Jones with a variety of illegal drugs from August 2000 through September 2001. Heclaims to have supplied her with a substance called "the clear," EPO, human growth hormone, andinsulin.

Reduce Fat / Cellulite

Gain Strength

Improve Memory

Re-Hydrate Skin

Regulate Sleep

Heighten Mood

And Much More...

Bovine growth hormoneInjection of recombinant BGH increases milk production and

is often used in dairy farming.

Pros:

Increased milk production.

Reducing the number of dairy cows needed reduces the

production of methane, a greenhouse gas.

Cons:

Concern about health risks of having BGH and additional

proteins in milk.

Increased mastitis (udder infection) in cows.

Agribusiness uses BGH to compete with small dairy farmers,

who can!t afford the drug.

Transgenic mice and fish

The growth hormone gene has

been engineered to be expressed

at high levels in animals.

The result--BIG ANIMALS

The rat GH gene was cloned behind the metallothionein

promoter that is induced with heavy metals. The mouse

on the left has the transgene and was fed heavy metals.

Transgenic mice are 2-3 times larger than siblings thatlack the transgene.

Transgenic mice

Ocean pout antifreeze gene promoter-chinook salmon GH

transgene in atlantic salmon

One major concern:

Transgenic salmon will escape from fisheries and breedwith strains in the wild. If the transgenic fish have a

mating advantage and are less fit, they could have anegative effect on the normal population.

Potential solutions to this problem:

Raise sterile fish.

Raise the fish inland away from coastal waters and thusbreeding populations of salmon.