Introduction to biotechnology- CLIL module for italian students by teacher Elena Bruno

1. Biotechnology
CLIL MODULE ABOUT…
Author: Prof.ssa Elena Bruno

2. UNIT 1
CONTENTS
• What’s biotechnology?
• How old is biotechnology?
• Differences between
traditional and
innovative biotechnology
• Recombinant DNA
technology
• Production of insulin
with recombinant DNA
technology

3. What is biotechnology?
Biotechnology is the use of living cells or organisms to produce materials
useful to people.
Traditional
Biotechnology
Innovative
Biotechnology
Use of living beings and selection of
natural features in living beings
use of genetically engineered
microorganisms, plant and animal cells
GMO

4. How Old Is Biotechnology?
10,000 BC
Domesticating Crops
1880’s Production of Vaccines
Domesticating Animals
8,000-9,000 BC
4,000 BC Leavening Bread
6,000 BC Brewing Beer
1940’s Production of Antibiotics
1980’s Use of genetically modified
organisms
molds
yeasts

5. Do traditional biotechnology
modify living beings?
Old biotechnology modifies living beings by selective breeding of
plants/animals: on the contrary of modern biotechnology, scientists do not
directly modify the DNA

6. Do traditional biotechnology
modify living beings?
The traditional biotechnology also use radiations (such as X-rays) to increase
the rate of mutations in the DNA of a species (such as wheat): in this way it is
possible to increase the genetic variability in a short time and to obtain
favorable variations. For example the variety "wheat Creso" was obtained in
this way.
irradiated wheat is not a GMO!

7. Do innovative biotechnology
modifies living beings?
Modern biotechnology modify
genetic material (DNA) in a way
that does not occur naturally by
mating and/or natural
recombination
EXAMPLE: Resistance against insects is achieved by incorporating into the food plant the gene
for toxin production from the bacterium Bacillus thuringiensis (Bt)
CORN COB

8. Applications of biotechnology
Medical Biotechnology
• Diagnostics
• Therapeutics
• Vaccines
Agricultural
Biotechnology
• Plant agriculture
• farms
• Food processing
Environmental
Biotechnology
• Cleaning through
bioremediation
• Preventing
environmental
problems
Forensic Biotechnology
• Paternity test
• Scientific
investigations
Industrial
Biotechnology
• Energy production
• New materials

9. APPLICATIONS OF BIOTECHNOLOGY

10. Some tools of biotechnology
Recombinant DNA technology
Gene Therapy
Cloning
Monoclonal Antibodies’
production
Stem cells
PCR
DNA fingerprint
DNA sequencing
In vitro fertilization
……AND MORE!!!!

11. Recombinant DNA technology
Recombinant DNA technology is one of the recent advances in
biotechnology, which was developed by two scientists named
Boyer and Cohen in 1973.
Recombinant DNA technology works by taking DNA from two
different sources and combining that DNA into a single molecule.
That alone, however, will not do much. Recombinant DNA
technology only becomes useful when that artificially-created
DNA is reproduced. This is known as DNA cloning.

12. What’s recombinant DNA?
RECOMBINANT DNA : is a DNA molecule that has been made
in the laboratory using at least two different sources of DNA
Recombinant DNA is produced using in vitro molecular
techniques to isolate and manipulate fragments of DNA
?
To produce Recombinat DNA you need
Molecular scissors:
Restriction enzyme
Molecular glue:
Ligasi enzyme
A vector:
Plasmide, viruses,
etc

13. Restriction enzymes
Enzymes that cleave DNA are used to manipulate and recombine
DNA.
Restriction enzymes, naturally used by bacteria as defense against
bacteriophages, cut DNA into smaller pieces.
Restriction enzymes cut DNA at
specific base sequences called
restriction site.
CUT=CLEAVE

14. Restriction enzymes
Some restriction
enzymes
recognise and
cut palindromic
DNA sequences
(sequences that
read the same
way in both
directions).
Some of them
make staggered
cuts, generating
fragments with
sticky ends

15. Restriction enzymes
Sticky ends

16. 16
Enzyme ligase
The sticky ends may join: a fragment from one source, such as human, can
be joined to a fragment from another source, such as bacterium. Initially
the fragments are held together only by weak hydrogen bonds: for these
reason you use the enzyme ligase to catalyze the formation of covalent
bonds between adjacent nucleotides at the ends of the fragments, joining
them to form a single, larger molecule
They join
ligase

17. 17
Enzyme ligase

18. 18
Plasmids
A plasmid is a small DNA molecule within a cell that is physically separated
from a chromosomal DNA and can replicate independently. They are most
commonly found in bacteria as small, circular, DNA molecules
• Plasmids are naturally exchanged among organisms
• They can be modified and used to deliver new pieces of DNA into an
organism=VECTOR
They often contain genes for antibiotic resistance
http://education-portal.com/academy/lesson/what-is-a-dna-plasmid-importance-to-genetic-engineering.html#lesson

19. Six steps of recombinant DNA
technology
1. Isolating (vector and
target gene)
2. Cutting (Cleavage)
3. Joining (Ligation)
6. Selecting (Screening)
5. Cloning
2
3
1
4
4. Transformation
(Introduction of the plasmid into
cells) 5
6

20. Six steps of recombinant DNA
technology

21. Screening
ampicilline resistance gene
lacZ gene
unfunctioning lacZ gene
human gene
Plate out the transformed
bacteria on a solid nutrient
medium containing ampicillin
and a sugar called X-gal.
Only transformed
bacteria that have
the ampicillin-
resistance plasmid
will grow.
This bacterium express lacZgene, will
hidrolyse sugar and form BLUE colony
This bacterium DOESN’T
express lacZgene, it isn’t
able to hidrolyse sugar,
will form WHITE colony:
it’s a GMO with useful
external gene
Repoter
genes or
marker
genes

22. Screening

23. Production of insulin in the past
Type I diabetes mellitus is a metabolic disease
caused by lack of hormone insulin.
It’s treatable only with injection
In the past, insulin was taken from the pancreases
of cows and pigs. The method was expensive and
some patients had severe allergic reaction to the
foreign insulin or its contaminants
these problems
have been
overcome thanks
to a GMO
http://education-portal.com/academy/lesson/what-is-genetic-engineering-definition-and-examples.html

24. 24
Production of insulin with
recombinat DNA technology
simplified
version

25. Production of insulin with
recombinat DNA technology
Correct
version
Two genes are chenically synthesized, one
for the insulin A chain and one for the
Insulin B chain.
Plasmid are extracted from Escherichia
coli. To make the bacteria produce insulin,
the insulin gene must be linked to the
beta-galactosidase gene, which carries a
promoter for tanscription.
The recombinant plasmids are inserted
back into bacteria, these bacteria
reproduce so that Insulin gene are cloned.
The GMO bacteria produce the two Insulin
chains that are extracted, purufied and
mixed together .
The A and B insulin chains conncet via
disulfade cross linkages to form the
functional insulin protein

26. 26
Animals are genetically engineered to exalt specific traits or
obtain useful products:
•GM animals can produce pharmaceuticals
•GM mice are bred for human disease research
•Xenotransplantation (animals as organs donor in
transplantation)
Genetically modified animals

27. Genetically modified animals

28. Genetically modified animals

29. UNIT 2
CONTENTS
• Biotecchnology in
medicine: production of
drugs
• Monoclonal antibodies
• Stem cells (blended
learning)
• Gene therapy

30. TPAis a naturally occuring human protein
that dissolves blood clots. It’used to treat
patients suffering from blood clotting in heart
attack or strokes
Production of drugs

31. 31
Production of drugs

32. Production of drugs with pharming
An expression vector
carrying a desired gene can
be put into an animal
fertilized egg (embryo),
which is implanted into a
surrogate mother. The
transgenic offspring
produce the new protein in
their milk. The milk is easily
harvested and the protein
isolated, purified, and made
clinically available to
patients.
Another way of making medically useful products in large amount is
pharming: the production of pharmaceuticals in farm animals or plants
2. The human
transgene is injected
into the Fertilized eggs
3. Eggs are
transferred to
recipient ewes
1. Donor ewes: after
insemination,
fertilized eggs are
collected
4. The offspring
are raised-
selection of
animals
producing the
human protein in
the milk
Ewe=female sheep

33. Production of drugs with pharming
• Human growth hormone (for
children suffering deficiencies) can
now be produced by transgenic
cows.
• Only 15 such cows are needed to
supply all the children in the world
suffering from this type of
dwarfism.
transgenic animals are not well seen by the public, and yet ....

34. Monoclonal antibodies
https://www.youtube.com/watch?v=Qp0z3bZDfKE

35. Use of Monoclonal antibodies

36. Use of Monoclonal antibodies

37. ADC: monoclonal antibodies drug
conjugated
https://www.youtube.com/watch?v=ACxmsMmo9hU

38. 38
Production of vaccines
Types of Vaccine
Whole-Agent Vaccine:
Contains whole, non virulent microorganisms
Subunit vaccine: Contains some part
or product of microrganisms that can
produce an immune response
Problems
These microrganisms can back-
mutate to a virulent form
Allergic reactions (e.g. against egg
substrate used to grow viruses)
They can be produced with
biotechnology, they are safer than
attenuated vaccines because they
can’t reproduce and they contain
little or no extra material (no allergic
reactions)

39. 39
Production of recombinant vaccines

40. 40
Production of vaccines

41. Gene Therapy
Gene therapy is an experimental technique for treating disease by altering the patient's
genetic material. Most often, gene therapy works by introducing a healthy copy of a
defective gene into the patient's cells.
Researchers must find safe, efficient vehicles--called vectors--to carry the modified
DNA to targeted cells. There are several vectors already in use, including chemical
solutions, synthetic fat molecules, and viruses that have been modified so that they
are harmless.

42. Gene Therapy
Gene therapy may be classified into the two following types, only one of which has
been used in humans:
Somatic gene therapy
As the name suggests, in somatic gene
therapy, the therapeutic genes are
transferred into the somatic cells (non sex-
cells), or body, of a patient. Any
modifications and effects will be restricted
to the individual patient only, and will not
be inherited by the patient's offspring or
later generations
Germline gene therapy
In germline gene therapy, germ
cells (sperm or eggs) are modified by the
introduction of functional genes, which
are integrated into their genomes. Germ
cells will combine to form a zygote which
will divide to produce all the other cells
in an organism and therefore if a germ
cell is genetically modified then all the
cells in the organism will contain the
modified gene. This would allow the
therapy to be heritable and passed on to
later generations
technical and ethical
problems=eugenics

43. Gene Therapy

44. Gene Therapy to treat SCID
A form of SCID is caused by a
deficiency of an enzyme
(adenosine deaminase or
ADA), which is necessary for
lymphocytes to develop.

45. http://learn.genetics.utah.edu/content/genetherapy/gtdoctor/
Space Doctor
Welcome to the Extraterrestrial Gene Therapy Lab! As Space Doctor,
you'll use your gene therapy knowledge to treat three ailing aliens who
come into your clinic.
A bit of fun…with gene therapy

46. Stem cells
https://www.youtube.com/watch?v=2-3J6JGN-_Y
ESL VIDEO: A Stem Cell Story Quick Quiz

47. UNIT 3
• Pro and cons of biotechnology

50. PRO: advantages of GMOs
Insect Resistance
Some GMO foods have been modified to make them more resistant to insect pests.
The University of California in San Diego reports that a toxic bacterium can be added
to crops to make them insect repellent, yet safe for human use. This can reduce the
amount of pesticide chemicals used on the plants, thus potentially reducing
exposure to pesticides.

51. PRO: advantages of GMOs
More Nutritious Foods
The Food and Agricultural Organization of the United Nations reports that some
GMO foods have been engineered to be more nutritious in terms of mineral or
vitamin content. Not only does this help you get the nutrients you need, it can also
play a significant role in battling malnutrition in the developing world. The United
Nations advises that vitamin A-enhanced rice is helping to reduce global vitamin A
deficiencies.
Golden rice

52. PRO: advantages of GMOs
More Nutritious Foods
Genetically-modified purple tomatoes : Their dark pigment is intended to give
tomatoes the same potential health benefits as fruit such as blueberries.

53. PRO: advantages of GMOs
More resistant plant
•Cold tolerance An antifreeze
gene from cold water fish has been
introduced into plants such as tobacco
and potato. With this antifreeze gene,
these plants are able to tolerate cold
temperatures that normally would kill
unmodified seedlings.
•Drought tolerance/salinity
tolerance Creating plants that can
withstand long periods of drought or
high salt content in soil and
groundwater will help people to grow
crops in formerly inhospitable places

54. tomatoes long conservation

55. 55
Edible vaccines
Plants enginereed to contain the vaccine can be grown locally, in the area where
vaccination in required, overcoming the logistic and economic problems of
transporting prepared vaccines over long distances. Most immportantly, edible
vaccines do not require syringes, saving money and eliminating the risk of
infection from contamined needles.
PRO: advantages of GMOs

56. 56
Bioremediation
PRO: advantages of GMOs
GM bacteria can clean up oil contamination repidly and effectly, but can you
think about the effect caused by the uncontrolled spread of these GM bacteria?

58. consumers are the guinea pigs?

60. CONS: problems with GMOs
Allergic Reactions
GMO foods can present significant allergy risks to people. Genetic modification often
mixes or adds proteins that weren't indigenous to the original plant or animal, causing
new allergic reactions in the human body. In some cases, proteins from an organism that
you're allergic to may be added to an organism that you weren't originally allergic to,
prompting the same allergic reaction experienced from the first organism.

61. CONS: problems with GMOs
Possible «escape» of transgene from crops to other species
Gene Transfer
A constant risk of GMO foods is that the modified genes of the organisms may escape into the
wild. Brown University warns that herbicide-resistant genes from commercial crops may cross
into the wild weed population, thus creating "superweeds" that are impossible to kill with
herbicides
More than ten
resistant weed
species have
appeared in the
U.S

62. CONS: problems with GMOs
Gene Transfer
A related risk is that the escape of genetically enhanced animals and vegetation can
create new super-organisms that can out-compete natural animal and plant
populations to drive certain species into extinction.

63. CONS: problems with GMOs
some crop are genetically modified to resist herbicides, this
encourages extensive use of herbicides. Another negative
impact is the possibility that increased us of an herbicide will
select for weeds with naturally mutations that make them
resistant to that herbicide (such as glycofosfate)

64. CONS: problems with GMOs
Economic concerns
Bringing a GM food to market is a lengthy and costly process, and of course agri-biotech
companies wish to ensure a profitable return on their investment. Many new plant genetic
engineering technologies and GM plants have been patented, and patent infringement is a
big concern of agribusiness. Yet consumer advocates are worried that patenting these new
plant varieties will raise the price of seeds so high that small farmers and third world
countries will not be able to afford seeds for GM crops, thus widening the gap between the
wealthy and the poor

65. CONS: problems with GMOs

66. CONS: problems with GMOs
Ethical issues
See-Through Frog.
GMO Bulls
GM animals may suffer poor health and reduced life span
Bald Chickens