Genetic Modification

& Biotechnology (3.5)

IB Diploma Biology

Essential Idea: Modern understandings of genetics and biochemistry

allow biologists to modify and manipulate the traits of organisms

Biologists have developed techniques for artificial manipulation of

DNA, cells and organisms
U 3.5.1 Gel electrophoresis is used to separate proteins or fragments of DNA
according to size and charge.

Gel Electrophoresis:
• Samples of either DNA or protein are
inserted into wells in a gel
• Gel is placed in a conducting fluid
and a current is passed through it
• Molecules move through gel based
on their charge (ex. DNA molecules
move to positive electrode since
they’re negatively-charged)
• Smaller fragments / molecules move
further through the pores in the gel,
while larger pieces don’t travel as far
U 3.5.2 PCR can be used to amplify small amounts of DNA.

The Polymerase Chain Reaction (PCR)

Synthetic method of amplifying specific
sequences of DNA. Useful when only a small
amount of DNA is available for testing e.g. crime
scene samples of blood, semen, hair, etc.

• Processes artificially recreates DNA replication

• Taq DNA Polymerase is used for PCR
• Comes from heat-resistant bacterium, Thermus
aquaticus, that lives in hot springs…
• Can resist denaturation at high temperatures
required to separate DNA strands in PCR
• Copies up to 1000 nucleotides / minute

http://highered.mcgraw-hill.com/olc/dl/1
20078/micro15.swf
U 3.5.2 PCR can be used to amplify small amounts of DNA.

The PCR Process:

PCR occurs in a thermal cycler and involves 3 steps:

1. Denaturation: DNA sample is heated to

95⁰C to break hydrogen bonds and separate it
into two strands

2. Annealing: DNA sample is cooled to 54 ⁰C,

allowing primers attach to opposite ends of the
target sequence

3. Elongation: A heat-tolerant DNA

polymerase (Taq) copies the strands

• One cycle of PCR yields two identical copies of

the DNA sequence
• A standard reaction of 30 cycles would yield
1,073,741,826 copies of DNA (230)
U 3.5.3 DNA profiling involves comparison of DNA.

http://learn.genetics.utah.edu/content/labs/gel/

http://www.dnalc.org/resources/animations/gelelectro
phoresis.html
U 3.5.3 DNA profiling involves comparison of DNA.
AHL: 7.1.8 Explain how tandem repeats are used in DNA profiling.

• Variable Number Tandem Repeats

(VNTRs) are short base sequences that
show variation between individuals in
terms of the number of repeats
• These are examples of Highly Repetitive
Sequences – useful for DNA profiling
due to uniqueness
APP 1 Use of DNA profiling in paternity and forensic investigations.

Forensic Investigations:
• Straightforward – just look
for a full match between
the DNA sample and the
potential suspects

Paternity Investigations:
• More complicated – since offspring inherits a mix of DNA from parents,
the child will show bands unique to each parent
• Each band in the child’s profile must match to a either a band in the
mother’s profile OR a band in the father’s profile (usually ~50-50 split)
SKILL 2. Analysis of examples of DNA profiles.
SKILL 2. Analysis of examples of DNA profiles.
SKILL 2 Analysis of examples of DNA profiles.
SKILL 2. Analysis of examples of DNA profiles.
SKIll 2 Analysis of examples of DNA profiles.
SKILL 2 Analysis of examples of DNA profiles.
SKILL 2 Analysis of examples of DNA profiles.
SKILL 2 Analysis of examples of DNA profiles.
SKILL 2. Analysis of examples of DNA profiles.
SKILL 2 Analysis of examples of DNA profiles.

Need to deduce whether or not a man

could be the father of a child from the
Biozone 132 - 135 pattern of bands on a DNA profile
U 3.5.4 Genetic modification is carried out by gene transfer between species.

Since the genetic code is universal,

genes can be transferred between
species and still allow for the same
polypeptide to be translated.

Gene transfer can be used to give

organisms new characteristics usually
only found in other species.

Left, a tobacco plant that has been

modified by the addition of a glowing
gene (for the enzyme Luciferase)
naturally found in fireflies.

These organisms are known as GMOs

or Transgenic organisms
U 3.5.4 Genetic modification is carried out by gene transfer between species.
U 3.5.4 Genetic modification is carried out by gene transfer between species.
APP 2. Gene transfer to bacteria with plasmids using restriction
endonucleases (enzymes) and DNA ligase
Transferring genes into bacteria, such as E. coli
requires plasmids, restriction endonucleases, and
DNA ligase enzymes
• An mRNA transcript encoding the desired
protein is obtained from the eukaryotic cell and
then made into cDNA (complimentary DNA
using reverse transcriptase)
• Not only is mRNA easier to extract (since
eukaryotic DNA is bound up with histones in the
nucleus), but this also ensures the inserted gene
will already be spliced and have no introns
(which bacteria do not have)

• A bacterial plasmid and the eukaryotic cDNA are

both cut with the same restriction enzymes
• DNA Ligase is used to seal the eukaryotic
sequence into the bacterial plasmid
APP 3. Assessment of the potential risks and benefits associated with genetic
modification of crops.
GMO Description
Rice modified with daffodil genes to have more
Golden Rice beta-carotene, which body converts to Vitamin A

Salt-
resistant Tomatoes modified to grow well in saline soils
Tomatoes

Corn modified with a bacterial insecticide gene so

Bt Corn
that it produces insect toxins within its cells

Sheep modified with human clotting factor IX gene

Factor IX
Sheep so that they produce clotting factor in their milk for
hemophiliacs

Soybeans modified with a herbicide resistance

Round Up
gene so farmers can spray fields and kill weeds, not
Ready Soy soybean plants

Rainbow Papaya modified with viral genes that make it

Papaya immune to the Papaya Ringspot Virus

Biozone 136
APP 3 Assessment of the potential risks and benefits associated with genetic
modification of crops.

Benefits of GMOs
Environmental Health Agricultural
Nutritional value of foods can Crops can be made to be
Pest-resistant crops mean be improved by enhancing drought, cold, and salinity-
vitamins
less chemical insecticides resistant, expanding range for
are used Crops can be produced that farming and increasing crop
yields
lack natural allergens or toxins
Less need to plow and
spray crops also save fuel, Herbicide resistant GM crops
GM crops can be engineered
reduced carbon footprint allow for easy killing of weeds
to produce cheap, edible that sap nutrients from crop
vaccines
Improved shelf-life plants
means less wasted /
GM bacteria produce cheap
spoiled food in stores Crop varieties can be produced
medical compounds such as
insulin and clotting factor that are resistant to viruses
APP 3. Assessment of the potential risks and benefits associated with genetic
modification of crops.

Risks of GMOs
Environmental Health Agricultural
Toxins in pest-resistant Proteins transcribed and GMOs with pest toxins could
GMOs could negatively translated from transferred increase evolution of
impact non-target genes could cause allergic
resistance in certain pest
organisms and harm reactions in humans or other
ecosystems animals – currently GM foods populations
are not necessarily labeled
Big biotech companies hold
Cross-species pollination monopolistic legal rights
could spread herbicide Antibiotic resistance genes
(patents) over GM seeds and
resistance genes and create used as markers during gene farmers must pay large sums
‘super-weeds’ transfer could spread to
for seeds each year. They are
pathogenic bacteria not permitted to save and re-
Biodiversity could be
sow seeds, so strains are not
negatively affected by Transferred genes could
destruction of pests, weeds, mutate and cause able to adapt to local
conditions.
and even competing plants unexpected risks
SKILL 3. Analysis of data on risks to monarch butterflies of Bt crops
Previously, farmers would protect crops
from pests by spraying with chemical
pesticides. Today, may crops are
genetically-modified with a gene from
the bacterium Bacillus thuringiensis (Bt)
that produces a protein toxic to insects

The Bt toxin kills targeted pest like the

corn-borer worm, but also kills non-
target insects

Monarch butterflies feed on milkweed

which often grow near Bt crops. When In the graph above, blue
pollen from the Bt crops ends up represents plants not
dusted with any pollen,
dusting milkweed plants, butterflies
yellow represents non-
consume the toxin and die
GM pollen dusting, and
red represents Bt pollen
dusted milkweed plants
U 3.5.5 Clones are groups of genetically identical organisms, derived from a
single original parent cell

Clone:
A group of genetically-identical organisms
derived from a single original parent cell

Organisms that reproduce asexually always

produce genetically-identical offspring (clones)

Clones are rarer in sexually-reproducing

organisms (i.e. monozygotic twins)

A clone can be very large, such as in the case of

commercially grown potatoes, but it can always
be traced back to an original parent cell.
U 3.5.6 Many plant species and some animal species have natural methods of
cloning

Many plants can naturally produce

clones (term derives from Greek
word for twig)

Examples:
• A single garlic bulb will clone itself to
form many identical bulbs in a
growing season

• Strawberry plants grow stems with

plantlets that can become
independent parent plants

• Hydra create clones by budding

SKILL 1. Design an experiment to assess one factor affecting the rooting of
stem cuttings

A stem cutting is a short length of plant stem that can be used to clone a plant. If
roots develop from the cut stem, it can become a new, independent parent plant

Nodes are parts of stem where leaves attach. Cuttings are made below nodes.
Normally takes several weeks for ‘rooting’ (new root growth)

Possible factors to study:

• Cutting above or below node
• Length of cutting
• Whether end is left in air or
Do this experiment and compost / water
use plants. Use plants • How many leaves are left on
• Use of hormone root powder
that form roots readily. • Type of compost
• Temperature
U 3.5.7 Animals can be cloned at the embryo stage by breaking up the
embryo into more than one group of cells
In early stages of embryo
development when cell are still
pluripotent, an embryo can be
fragmented or split to create animal
clones. This is how monozygotic twins
can occur in humans.

Animal embryos created through in

vitro fertilization can be artificially
fragmented and then transplanted
into surrogate mothers. This is most
effectively done when the embryo is at
the 8-cell stage

Less interest in this cloning process since the 8-cell embryo

stage is still too early to assess if a clone will have desired
traits or not…
U 3.5.8 Methods have been developed for cloning adult animals using
differentiated cells
In the 1950s, John Gurdon, then a
student at Oxford, removed the
nucleus from the body cell of a
tadpole and transplanted the nucleus
into a tadpole egg cell.

The egg cell with the transplanted

nucleus developed as a normal zygote
and created a tadpole with the same
genome as the body cell nucleus

In 1996, Dolly the sheep became the

first mammal cloned in this way

In 2012, Gurdon received the Nobel

Prize in Medicine for his research
APP 4. Production of cloned embryos by somatic cell nuclear transfer

Cloned embryos, such as Dolly, are

produced by a process called Somatic
Cell Nuclear Transfer (SCNT)
1. Somatic cells are taken from adult organism to
be cloned and grown in a low-nutrient medium.
This inactivates genes to wipe out any previous
pattern of differentiation
2. Unfertilized egg cells are taken from a female of
same species and their nuclei are removed
3. Cultured somatic cells and enucleated egg cells
placed side-by-side and zapped with a small
electric pulse to fuse them together (about 10%
of cells fuse)
4. Fused egg cells containing somatic cell nucleus
develop into embryos for seven days and are https://www.hhmi.org/biointeractive/somatic-cel
l-nuclear-transfer-animation
then implanted into surrogate mother (in the
case of Dolly, only 1 of 29 successfully implanted
and completely developed)
APP 4. Production of cloned embryos by somatic cell nuclear transfer
APP 4 Production of cloned embryos by somatic cell nuclear transfer
Bibliography / Acknowledgments

Bob Smullen