AGRIC 3120 – BASIC BIOTECHNOLOGY
------Part 1
 BIOTECHNOLOGY
- is the manipulation of natural biological processes in
order to serve societal needs.
 Four main areas of Biotechnology
 Transgenic Biotechnology - Mixing genetic
material from multiple sources (species)
 Reproductive cloning - Techniques used to clone
certain species (mammals)
 Reprogramming of Cells - Reprogramming
differentiated cells or using stem cells to become
needed tissues in patients with diseases or physical
harm
 Forensic Biotechnology - Use of restriction
enzymes and electrophoresis to distinguish one
person from another
 Transgenic Biotechnology
 HGH Deficiencies
The pituitary gland produces a crucial hormone called
the human growth hormone.
• This peptide hormone (protein) provides for normal
growth and development.
• If the pituitary gland is defective then growth is
severely stunted.
• For many years HGH had to be extracted from the
pituitary glands of deceased humans which meant that
there was a shortage of available HGH.
Now, we have all we need! How?
• The HGH gene was cut out of the human genome and
inserted into a plasmid, which is now now called
recombinant DNA because it contains DNA from
multiple sources.
• The plasmid is then taken up via transformation by a
bacterium.
• The bacterium reproduces many timesand when the
the bacterium undergoes transcription and translation
(protein synthesis), it makes all of the HGH that we
could possibly need!
 PLASMID - is a small, circular piece of DNA that
not only is separate from the chromosome, but can
also replicate independently.
•How do they cut the gene of interest out of the
genome?
- RESTRICTION ENZYME
How does a fragment then get spliced in?
 Insulin
The pancreas, among other functions, produces a
crucial hormone called insulin.
• This peptide hormone (protein) ensures that glucose
is taken up by the cells for cellular respiration.
• If the pancreas is defective then the blood sugar levels
get dangerously high causing many physiological
effects (Diabetes mellitus).
• Using very similar technique as HGH production
previously mentioned, scientists were able to use E. coli
to bioengineer synthetic insulin in 1977.
• Other transgenic organisms used to produce insulin
today are yeast (Saccharomyces cerevisiae) and a
plant called safflower (Carthamus tinctorius).
 Golden Rice
The World Heath Organization estimates that between
1 and 2 million children die each year from vitamin A
deficiency.
• Golden rice is a genetically modified food that is
fortified with beta carotene, which the human body
converts into vitamin A.
• This transgenic organism is the result of mixing genes
from a bacterium and from daffodils into the rice
genome.
• It is not currently used due to regulatory issues.
 Reproductive Cloning
 CLONE - It is an exact genetic replica of another
cell or organism.
What have we cloned so far?
• DNA (Polymerase Chain Reaction)
• Cells (creating tissue cultures or stem cell lines)
• Whole organisms
 Organismal Cloning
• What has been cloned thus far?
• Plants have been cloned for thousands of years!
• Bananas, potatoes, grape vines (grafting), etc.
• Many trees, shrubs, and vines are just clonal
colonies.
• Animals
• Parthenogenesis – development of a new
individual from an unfertilized gamete; aka unisexual
reproduction (sharks, anteaters, some insects, etc.)
• Some animals have undergone somatic cell
nuclear transfer such as: sheep, rats, cats, goats, dogs,
camels, and many others.
Somatic Cell Nuclear Transfer (SCNT)
So, what if we...
• What if we manipulate animal embryos and use
recombinant technology to give these animals some
beneficial characteristics ...to us? That is what some
scientists have been able to do
• Some animals, like this goat, have been bred to
produce certain peptide hormones needed by humans
when they express milk
. These proteins can easily be separated from the milk
for human use!
 Reprogramming cells  iPS cells
What can stem cell research do for us? • In 2007, the induced pluripotent stem (iPS) cells were
developed.
 Stem cells can help us in many medical • Reprogramming genes are spliced into normal human
applications such as: somatic cells.
• Organ and tissue regeneration • This tricks the cell into changing from a differentiated
• Fighting the following diseases: cell into a pluripotent cell.
• Cardiovascular disease • The cell can then develop into a desired, differentiated
• Brain diseases like Parkinson’s and Alzheimer's cell of another type!
• Blood diseases like leukemia and sickle-cell anemia
So...what’s all the fuss about? • THIS COULD ELIMINATE THE NEED FOR
The stems cells that work the best come from embryos. EMBRYONIC STEM CELLS!
 STEM CELLS - are undifferentiated, meaning that
they haven’t become a type of cell yet.
• When a sperm meets an egg, the resulting zygote is
totipotent. The inner cell mass, the source of
“embryonic stem” cells, are pluripotent.
• Totipotent cells have the ability to create a whole
organism, or at least all different types of tissues.
• Pluripotent cells can only give rise to most types of
tissues, and definitely NOT a whole organism.
Plant cells are totipotent!
 The 2012 Nobel Prize in Physiology and
Medicine
JOHN GURDON & SHINYA YAMANAKA
- They received the 2012 Nobel Prize (Physiology and
Medicine) for their work with the development of iPS
cells.
 Forensic Biotechnology
- is used to determine the identity of certain individuals:
• Criminals
• Disaster victims
• Biological parents
 PCR: Polymerase Chain Reaction
• Usually there is only a small amount of DNA to work
with at a crime scene.
• Investigators and forensic scientists use the
polymerase chain reaction to make thousands of
copies of key regions of the original DNA strand.
 Electrophoresis
• The PCR products (DNA strands) are analyzed via
electrophoresis for STR’s (short tandem repeats).
• Every person has their own individual pattern of these
STRs.
• For a single set of primers, a person will have 2 PCR
products if they inherited different numbers of STRs
from each parent. This results in 2 bands on their gel.
 Where is your line?
Other than the ones already mentioned, here are some
other “real-life” examples of biotechnology. Do any of
these cross your line?
• Injecting human brain cells into monkey brains
- for brain disease research
• Xenotransplantation
- using animal “parts” for our parts (for instance using a
pig valve to replace a defective heart valve in a human)
• Adding human stem cells to sheep fetuses
- to produce sheep with livers made of mostly human
tissue
• Bt crops • Pigs (PPL Therapeutics- delete a gene which causes
- these crops contain genes from the bacterium Bacillus hyperacute rejection of pig-to-huma n organ
thuringiensis which produces proteins toxic to pest transplantation)
insects • Primates (HIV and AIDs research, geriatric research)
• Roundup Ready crops
- contain genes that protect them from Roundup  Alternatives to Animal Models
(herbicide) • Cell culture devices
• Researchers use cell cultures and computer-
generated models whenever possible, but this doesn’t
work for looking at an organ or entire animal
AGRIC 3120 – BASIC BIOTECHNOLOGY
------Part 2  Regulation of Animal Research The “Three Rs”
• Reduce the number of higher species (cats, dogs,
 Animals in Research primates) used
• Replace animals with alternative models whe never
possible
• Refine tests and experiments to ensure the most
humane conditions possible
 Veterinary Medicine as Clinical Trials
• Treatments for humans may also be useful for
treatments with animals (e.g. the BRCA1 gene found in
65% of human breast tumors is similar to the BRCA1
gene in dogs)
• Hyperthermia + radiation = more effective at killing
tumors
• Stimulation of cytokines for curing skin cancers
 Bioengineering Mosquitoes to Prevent Malaria
❑ Cloned a gene that prevents the parasite from
traversing the midgut; blocking the continuation of its
life cycle
 Animal Models
• Mice ❑ Developed an antibody that prevents the parasite
• Rats from entering the mosquito’s salivary gland
• Zebrafish (3 month generation time, 200 progeny,
complete embryogenesis in 120 hrs)  Cloning
• Dogs (lungs and cardiovascular system) • Embryogenesis – the process by which the embryo
• Cats forms and develops
• Zygote – fertilized egg
• Blastocyst – early stage embryo prior to implantation  Transgenic Techniques
 Embryonic Stem Cell Method
 Cloning Methods  Pronuclear Microinjection
❖ Embryo Twinning  Making clones of a transgenic animal
 Transgenic Applications
❖ Increased Production Efficiency: Transgenic Growth
Hormones
❖ Improved Food Safety and Quality: Longer Shelf Life
❖ Improved Food Safety and Quality: Lactose
Intolerance
❖ Increased Production Efficiency: Boost Lactational
Performance
❖ Disease Resistant Animals less susceptible to
mastitis
❖ Disease Resistant Animals less susceptible to mad
cow disease
❖ Somatic Cell Nuclear Transfer ❖Decreased Environmental Impact
AGRIC 3120 – BASIC BIOTECHNOLOGY
------Part 3
 What is the Result of Biotechnology?
- An organism showing a novel trait not normally
found in the species
 Limits to Cloning
 Decrease Genetic Diversity
 Efficiency and Cost Effectivenes
 Abnormal Development
 Premature Aging  Biotechnology Terms You Probably Heard
 The Future of Cloning  Transgene – the foreign gene added to a species
 Increase in genetic gain Ex. – modified EPSP synthase gene (encodes a protein
 Consistent Quality that functions even when plant treated with Roundup)
 Endangered Species (Adult Gaur & Noah the  Transgenic – an organism containing a transgene
cloned Gaur) introduced by technological (not breeding) methods
Ex. – Roundup Ready Crops
 Biotechnology Develops
 GMOs - Genetically modified organisms
• GMO - an organism that expresses traits that result
from the introduction of foreign DNA
• Also called transgenic organism
 Important Terms
Source: USDA
 Breeding
✓Beneficial gene added from the same species
✓Gene delivered by mating within the
Species
 Transformation
✓Beneficial gene added from another species
✓Gene delivered by plant genetic trans formation/
engineering
 Let’s Be Up Front
• Breeding =/Biotechnology
✓ Breeding only exchanges genes found in the species
✓ Breeding can transfer the transgene to other
breeding materials
✓ BUT it is not the same as biotechnology
• Biotechnology adds traits not available in the
species
✓ Soybean does not have a gene to breakdown
Roundup
✓ The gene comes from bacteria
 Interspecific Cross
 Mutagenesis: New Trait, No Foreign Gene
✓ Mutagenesis changes the sequence of a gene
✓ New, useful traits can be obtained

 BASF Clearfield Products

 Mutagenesis Crops

 Herbicide resistance
• Imidazolinone herbicides, which include imazapyr,
imazapic, imazethapyr, imazamox, imazamethabenz
and imazaquin, control weeds by inhibiting the enzyme
acetohydroxyacid syn thase (AHAS)
 Mutant AHAS enzyme
• Developed by mutagenesis
Crops
• Canola, Corn, Rice, Sunflower, Wheat
 In US
• Not considered GMOs by USDA regulators
• A Major marketing advantage

Note:
Some of these herbicide resistant genes have been
stacked with transgenes, so those products are also
considered to be transgenics or GMOs.