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.
Salient Features of India constitution especially power and functions
Application of recombinant dna in the pharmaceutical and medical fields
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
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
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.
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
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
31.
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