Gene and Protein Engineering in Clinical Medicine Flashcards
Production of Proteins for Use in Treatment of Disease The following goals must be achieved?
- Isolation of the gene (cDNA)
- Introduction of the gene into a host cell system where
it is expressed - Growth of host cells on a commercial scale 4. Purification of product to meet rigorous standards
- Extensive testing to establish clinical efficacy
- Approval from regulatory authorities for clinical use
Example of host cells ?
➢ Bacteria, yeast or
➢ Cultured insect or mammalian cells
Choice of host cells depends on what?
And what is the difference between Mammalian ,Bacteria and yeast ?
Choice depends largely on properties of the
protein and particular features of each host cell system.
Bacteria and yeast are easier to grow than
mammalian cells.
• Mammalian cells handle expression of
larger secreted proteins better than bacteria
or yeast.
What Medically Useful Proteins
?
Recombinant DNA and expression vectors
have been used to make medically useful proteins that would otherwise have been difficult to obtain in necessary quantities.
What is the TPA ?
produced by cells lining the blood vessels (enzyme)
converts plasminogen, found
normally in the blood, into plasmin, a protein
that dissolves clots.
What is the Human Growth Hormone ?
*Synthesized and secreted by the pituitary at the base
of the brain
• Regulates of growth and development
HGH is species specific; only the human
protein works.
If there is HGH deficiency what happen ?
hypopituitary dwarfism Must treat early while bone
growth is possible.
Viral Disease Therapy example ?
Hepatitis B
DNA Vaccines
Synthesise of the viral protein for Hepatitis B
successful in ?.
Synthesise of the viral protein; failed in
bacteria, but was successful in yeast!
Hepatitis B vaccine Tested for effectiveness in ?
primates and then
in human volunteers.
Hepatitis B vaccine first use in ?
Australia
Present gem of efficacy on Hepatitis B vaccine.
94.5% efficacy
Hepatitis B vaccine can be effective only if ?
takes place before
infection
Hepatitis B vaccine
And DNA Vaccines
Based on what ?
Hepatitis B vaccine
• Based on recombinant HBSAg.
DNA Vaccines
• Are based on use of plasmids containing
protein coding genes.
How can we identify plasmids key to defending
against microbial pathogens ?
• Cut pathogen DNA into fragments and clone
into a plasmid to make a library.
• Divide the library into pools
. • Test pools in animals; give time to develop
response then inject pathogen.
Establishing New DNA Vaccines
▪ Assume any unaffected animal group contains
at least one “protective” plasmid from a pool. ▪ Subdivide this pool and repeat the testing cycle
until a single/few plasmids are identified that
confer protection.
Some medically useful products of biotechnology ?
Table 18.1
Plant Biotechnology Applications targeting what?
nutrition and the
environment.
β-carotene is converted to what in our body ?
vitamin A
Example of Adapting A Crop Plant To An Environment?
plants that are salt-tolerant
They take the gene for enzymes that synthesize β-carotene
from the precursor are taken from??
daffodils
They take the gene that responsible of protein that moves sodium ions into
the central vacuole were isolated from??
Arabidopsis
Concerns Over Plant Biotechnology.?
• Genetic manipulation is an unnatural
interference in nature.
• Genetically altered foods are unsafe to eat
. • Genetically altered crop plants are dangerous
to the environment.
➢ Escape of transgenes into wild populations: ➢ if the gene for herbicide resistance made its way
into weeds
. ➢ beneficial insects killed from eating plants with B.
thuringiensis genes.
Hepatitis B immunisation requires how many injections?
3 injections
over a 6 month period
Which Vaccine did the lettuce made a and which proteins
Measles Vaccine
By Measles virus hemagglutinin (MV-H) protein
Advantages of lettuce Measles Vaccine
.
Offers development of measles vaccine formulation that
is effective, temperature-stable, easy to administer in a
resource-poor setting and amenable to large scale
manufacture
Example of
use transgenic dairy animals
that produce useful products in their milk.
-1AT in humans inhibits what ?
elastase, which
hydrolyzes connective tissue.
If there-alpha -1AT deficiency’s what will happen ?
Excess elastase is found on the surface of the
lungs of people with cystic fibrosis and is
partially responsible for their severe breathing
problems.
Pompe’s disease ?. characteristic. Cause of disease .treatment?
diagnosed by absence of alight-glucosidase activity
Cardiac hypertrophy
Loss of muscle strength prevents infants from achieving developmental milestones such as sitting, standing, and walking
enzyme alpha glucosidase from the milk of transgenic
rabbits has been successfully used for treating infants
Why’s to block Translation of mRNA?
Using Antisense RNA and siRNA
