This document discusses the application of recombinant DNA technology in pharmaceutical and medical fields. It begins with an introduction and objectives. It then describes the major steps of genetic engineering which include isolation, cutting, ligation, transformation, selection, and expression of cloned DNA. Key applications discussed include producing human insulin through recombinant E. coli, treating growth hormone deficiency with recombinant human growth hormone, and developing vaccines by cloning antigen genes. The document concludes by stating recombinant DNA technology produces medical products that play an important role in treating genetic diseases and developing targeted medicines.

1. ekelle university
f natural and computational science
MAIREG
KEBEDE
Application of Recombinant DNA in the
Pharmaceutical and Medical Fields
COMPAILED
BY:-
UNIVERSITY OF
GONDAR
INSTITUTE OF BIOTECHNOLOGY
Advisor
Name:-
Pro. NEGA BERHANE (PhD)
Institute of Biotechnology,Director
General

2. BY: MAIREG KEBEDE 2

3. © Introduction
© Objectives of the Presentation
© Steps of Genetic Engineering
© Application of Recombinant DNA Technology in Pharmaceutical and Medical Fields
 Human Insulin
 Human Growth Hormone
 Vaccines
© Conclusion
BY: MAIREG KEBEDE 3

4.  At the end of this presentation we will be able to:
 Define Genetic engineering and Recombinant DNA technology.
 List down the six major steps of Genetic engineering.
 Briefly describe the application of Recombinant DNA technology in Pharmaceutical and
Medicine.
BY: MAIREG KEBEDE 4

5.  Genetic engineering or recombinant DNA technology is the procedures by which
DNA from different species can be isolated, cut and spliced together .
 New "recombinant " molecules are then multiplied in quantity in populations of
rapidly dividing cells (e.g. Bacteria, yeast).
 It is simply the addition, deletion, or manipulation of a single trait in an organism
to create a desired change.

6. 1. Isolation
1 (a) Isolation of a specific gene from donor e.g. human
 Cells broken open
 Genetic probe added
 Reveals position of the gene of interest
6BY: MAIREG KEBEDE
Position of
gene of
interest
Genetic probe
Donor DNA

7. (b) Isolation of plasmid from a bacterial cell
7BY: MAIREG KEBEDE
Bacterial cell
Plasmid

8. 2. Cutting
 Restriction enzymes act as molecular scissors and cut DNA at specific sites called
restriction sites.
BY: MAIREG KEBEDE 8
Restriction site
Restriction site
Restriction enzymes

9. BY: MAIREG KEBEDE 9
Restriction site Restriction site
Donor DNA
PLASMID
Restriction enzymes
© Biology Support Service 2007

10. BY: MAIREG KEBEDE 10
Donor DNA
Sticky EndsPlasmid

11. 3. Ligation
 Re-joining cut fragments of DNA and forming artificial recombinant molecules.
BY: MAIREG KEBEDE 11

12. 4. Transformation
 Recombinant DNA introduced into bacterial cell
BY: MAIREG KEBEDE 12
Bacterial
chromosome
Bacterial
cell
Recombinant DNA

13. 5. Recombinant selection or Screening
BY: MAIREG KEBEDE 13

14. 6. Expression of cloned DNA
 Reasons for attempting to express cloned genes
(i) The construction of expression libraries (cDNA libraries where the donor DNA of
each clone is expressed, allowing screening for structural or functional properties of
the encoded polypeptide)
(ii) The analysis or exploitation of gene function at the protein level;
(iii) The commercial production of proteins;
(iv) The production of antibodies
BY: MAIREG KEBEDE 14

15.  Recombinant DNA technology has made it possible to treat many diseases by replacing
damaged and diseased genes in the body with new genes.
 It has brought revolutionary changes in the field of medicine and introduced such methods of
treating diseases and delivering drugs that were once just imaginary.
 Human Insulin
 Human Growth Hormones we will disscussed briefly on those topics
 Vaccines
 Monoclonal Antibodies
 Interferon
 Antibiotics
BY: MAIREG KEBEDE 15

16. 1. Human Insulin
 Insulin is basically a hormone, which is made up of proteins. This hormone is
secreted in cells of the pancreas that are commonly referred to as the ‘Islets Of
Langerhans’.
 This hormone plays important role in controlling the glucose level in body, because
decreased level of insulin may cause diabetes.
 Recombinant DNA technology has allowed the scientists to develop human insulin
by using the bacteria as a host cell.
 A variety of different recombinant insulin preparations are in widespread use.
 Recombinant insulin is synthesized by inserting the human insulin gene into E.
coli, which then produces insulin for human use. This is supposed to be safer than
traditionally prepared drugs. 16

17.  It is a necessary hormone that enables bodily cells to allow blood sugar to enter and be
converted into energy.
 Pig and cattle pancreas glands were once the only viable method. This was sufficient for most
diabetics however there was issues:
 Animals insulin caused some allergic reactions.
 Not a true human match to insulin.
 Not the most efficient way to obtain insulin.

18.  Before insulin was discovered in 1921, everyone with diabetes died within weeks
to years of its onset.

19.  In 1969, Dorothy determined the spatial conformation of the molecule, the so-called tertiary structure.
She had been awarded a Nobel Prize in Chemistry in 1964 for the development of crystallography.
 In 1958 british molecular biologist Frederick Sanger determined the primary structure of insulin. It
was the first protein to have its sequence be determined. He was awarded the Nobel Prize in Chemistry
for his work.
 In 1977 Rosalyn received the Nobel Prize in Medicine for the develpoment of radioimmunoassay for
insulin.

20.  Synthetic insulin was first made in 1978 by scientists at Genetech, Inc. and City of
Hope National Medical Center.
 Herbert Boyer was the scientist first who succeded in making a synthetic “human”
insulin through gene-engeneering in a laboratory in 1977.
 Made possible by the discovery of restriction enzymes & DNA ligase.
– Found naturally in bacteria.
– Catalysts to cut and rejoin DNA fragments.

21. BY: MAIREG KEBEDE 21
Human Insulin Production by Bacteria

22. BY: MAIREG KEBEDE 22

23. BY: MAIREG KEBEDE 23

24. BENEFITS
 Less expensive
 Absorbed more rapidly by the body
 Has a shorter more manageable duration of effectiveness
 Causes fewer allergic or autoimmune reactions than the animal insulin hormone.
SIDE EFFECTS
 Extreme lethargy
 Mental confusion
 Memory loss
 Joint and muscle pains
 Depression
 General feeling of being unwell.
BY: MAIREG KEBEDE 24

25. 2. Human Growth Hormones
 Human growth hormone is a polypeptide hormone. It is responsible for growth,
reproduction of the cells and regeneration in humans as well as animals.
 It is secreted by somatotroph cells present in the pituitary glands. Before
recombinant HGH became available, HGH for therapeutic use was obtained from
pituitary glands of cadavers. This unsafe practice led to some patients developing
Creutzfeldt_x0002_Jacob disease.
 Recombinant HGH eliminated this problem, and is now used therapeutically. It has
also been misused as a performance enhancing drug by athletes and others.
 In recent days biotechnology has helped scientists to produce many growth
hormones. The dwarfism disease is successfully treated with this hormone
BY: MAIREG KEBEDE 25

26. Production of Growth Hormone
BY: MAIREG KEBEDE 26

27. How HGH methods work
 Injection: a synthetic substance injected into the body. They tend to be very expensive a
single injections costs as much as $29.
 Oral Spray: not scientifically proven to have an effect. Some say they see results faster then
the other methods.
 Natural releasers: they increase the HGH levels in our bodies. They are like vitamins taken
in a tablet form. Usually taken just before bed seeing how the pituitary gland is most active
during sleep.

28. Intended use
 Children with growth disorders
 Adults with hormone deficiency
 Short bowel syndrome(intestinal disorder)
Side effects
 Increased risk of diabetes
 High blood pressure
 Arthritis
 Joint swelling
 Joint pain
 Headache
 Nausea, vomititing
BY: MAIREG KEBEDE 28

29. 3. Vaccines
 Vaccine is a biological substance that is prepared from the suspension of weak or dead
pathogenic cells.
 It is injected in the body to enhance the production of antibodies against a particular
antigen.
 Recombinant DNA technology has made it easier for scientists to develop vaccines by
cloning the gene used for protective antigen protein.

30.  Edward Jenner used the cowpox virus to vaccinate individuals against smallpox
virus in 1796.
BY: MAIREG KEBEDE 30

32. DNA vaccines Vs Traditional vaccines
DNA Vaccines Traditional Vaccines
BY: MAIREG KEBEDE 32
 Uses only the DNA from
infectious organisms.
 Avoid the risk of using actual
infectious organism.
 Provide both Humoral & Cell
mediated immunity
 Refrigeration is not required
 Uses weakened or killed form of
infectious organism.
 Create possible risk of the vaccine
being fatal.
 Provide primarily Humoral immunity
 Usually requires Refrigeration.

33. ADVANTAGES
 Elicit both Humoral & cell mediated immunity
 Long term immunity
 Refrigeration is not required
 Stable for storage
DISADVANTAGES
 Extended immuno-stimulation leads to chronic inflammation
 Some antigen require processing which sometimes does not occur
BY: MAIREG KEBEDE 33

34.  Recombinant proteins are widely used as reagents in laboratory experiments and
to generate antibody probes for examining protein synthesis within cells and
organisms.
 Thus the use of this advanced technology, Recombinant DNA technology produces
variety of products which are used for medical purposes.
 It is a challenging field, and play a key role in preventing genetic diseases,
producing targeted medicines, and providing patients with less toxic
pharmaceuticals.
 Hence it is gaining tremendous significance in the field of medicine today.
BY: MAIREG KEBEDE 34