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Biotechnology II wild animal for meat ,dairy,liether,transport
Domesticated animals fewer than 20Cattle ,sheep , goast , pigs , hoarse. 5 species .
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Dosile
Flexible in their dietary
Can grow in road maternity quickly n a herbivore diet.
Breed readily n captivity.
Have hierarchical social structurers that permit human to established.
Dominance over them.
Adapted to living in large groups.____________________________________________________________________________
Development of farm animals
Selection methods .
Potential agricultural value.
The animals adapt to thrive in a domesticated environment.
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Vacccination
Freezing of semen.Testing n selection.
Use of antibiotic in feed to increase gain.
Embryo transfer.
Embryo splitting n cloning from blastomeres.
In vitro maturation in vitro fertilization of oocyte n in vitro resulting embryo.use of
hormones.
Hormone sex reversal.
Steroid administration to improve weight gain . Bovine somatotropin BST.
_____________________________________________________________________________Making transgenic animals.
Inject gene and feede fertilized egg of the desired animal using a very small glass
Needle visualized microinjection.
In approximately 1% of the injected eggs the gene is physically recombine an
eggs genome.
The new gene integrate into the genome before egg begin to ddivide.
Implanted the fertilized egg is into a mother where it fully develops transgenicanimal.
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Human disease research.
Input , output , value-added traits.
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Input traits
- Increasing production efficiency.
- More efficient allergics leaks growth rates.
- Resistance to disease._______________________________________________________________________
Output traits.
Enhancing the quality of the animal product.
eaner more ee an por .
Milk leaks allergics protein
,yogurt.
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Value-added traits
Therapeutics proteinsXenotransplantation.
Models for hujman disease.
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Biot in animal medication vaccine .
Medicine.
Production
Anti-bacterial compound and substitute using same antibiotion in animals.
Animal vaccine.
Rapid test kits.
_________________________________________________________________________Therapeutic in value.
Antithrombin III
Milk protein.
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Vector used for gene transferre escape n hazrd.
Environment issue.
Animal welfare issue.
The tool government department and agencies involved sufficiently well defined
n weather sufficient to cope with technologies.
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Regulation agency in USA.
FDA department of agriculture,environment protection agency to regulatetransgenic animals n their prodcuts to ensure that they are safe for public
uses and environment.
Depending on the nature of the genetic modification .
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Genetically identical copies___________________________________________________________________________
Monozygotic twins
Transfer to nucleated oocyte of nuclei from blastomere,blastomere
Nucleus transfer BNT.
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Soamtic cell nuclear transfer SNT
Embryonic stem cells.
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BNT,SNT result in an exact replica of an individual animal.Mitochondria ,cytoplasmic factors. Mitochondria ,cytoplasmic factors.
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Health problem.
Age prematurely.
Lack of genetic diversity ,
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Technical and application .
Germ line how transgenic animal produced?
DNA microinjection.Retrovirus mediated gene transfer.
Embryonic stem cell mediated gene transfer.
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,
their offspringAcquire the desired transgene.
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Male transgenic low
Gene transfer 0.6% transgenic pigs were injected with specific transgene.
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DNA microinjection
Transfer of the desired gene. Pronucleus.
The manipulate cell must be culture in vitro to develop a specific embryonicphase transfer to the recipient female.
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Retrovirus mediated gene transfer
Use vectors to transfer,resulting in a chimera and organism consisting
Of tissue , part of diverse genetic constitution.
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Chimera 20 generations.
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Embryonic stem cell mediated gene transferIsolation of totipotent stem cells.
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Agricultural
Breeding
Quality
Growth hormones.
Disease resistance.
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Medical transplantation.
XenotransplantationTransgenic pigs then transplant organ.
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Nutritional supplement and pharmaceuticals products ,insulin , GH ,anti-clotting
factors from milk transgenic from cows sheep goats
Treat PKU phenylketonuria ,cystic fibrocis.Protein-enriched milk 2.4 gram/litre
Rosies milk alpha lactalbumin.
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uman gene erapy
Normal copy of a gene (transgene) to the genome of a person defective copies ofthe gene.
- Genetic discover A.I. produce calf and gene then promoters the growth of red
cells in human.
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Industrial application.
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Industrial application.
Toxicity-sensitive produced for chemical safety testing.Microorganism engineered to rpoduced a wide variety of proteins then enzyme
speed up industry chemical reaction.
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Transgenic chicken.
Grow faster than sheep and goats .Synthesize several group of protein in the white of their eggs.
Chicken carrying an expressing foreign genes.
Infecting embryo with a viral vector copying.
uman gene or erapeu c pro e n .
Promoter sequences that will respond to the signals for making proteins(lysozyme) in egg white.
Transforming sperm and human gene and promoters checking transgenic
offspring.
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Transgenic fish
Design and construction of trasgenic DNA .Transfer gene to genome cells.
Screening for transgenic fish.
Find transgene expression and phenotype.
Study inheritance
Stable lines of transgenics.
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GH gene from large sized fish.
Human benefit protein.
Gene were introduced into medaka fish to create resistance to bacterial
pathogen.
Some commercially desirable fish and crutaceaus have been difficult to
genetically engineer because embryonic tissue is difficult to manipulate.
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Regulation of transgenic fish.
Synthetic triploid strain of fish eggs roduced from treatment of pressure
temperature shock and sex hormone the sterile triploids were tend toproduced fertile progeny established transgenic population.
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The tiny zebra fish that live in aquariums a popular laboratory animals to
produce a fluorescent red pigments.
,promoted for sale household aquarium.____________________________________________________________________________
FDA position glofish = unmodified fish.
Food safety isomers.
ene sease res s ance s o a sor ox c su s ance g.
90% food allergies ,potential for allergic reactions. Large dose it consume in
cow and uncook food
Suchi.
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Eu concerns
Natural ecosystem and native species
50% of all harmful economics and environment.
66% unintentionally.
Farmed fish excape from open water contaminate native population.
Inevitable that trangenic fish will escape from aquaculture .
Inevitable that transgenic fish.Competitive advantager over native species.
Using GH gram increase growth 2-11 times.
Lead to sexual maturity, breeding,
pee roug ou na ve popu a on.
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Maturity,factor,die sooner,goes for rapid growth,shorter life expectancy.
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Transgenic cow
1. Superovulation treated
2. Fertilization
3. Gene transform by microinjection.
4. Implantation.
5. Determination of transgene expression and phenotype.
6. Study of inheritance.7. Selection of stable lines of transgenic.
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Pharmaceutical.
Disease. Resistance.
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30% from staphylococus aureus then secreting added protein. Lysostaphin
Defense bioengineering.
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Transgenic cow and blood thickener in milk.
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Transgenic 3 years develop drugs.
Developed from ovaries of dead cow.
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Creat bioreactor for biopharnmaceuticals production of recombinant human growth hGH in the milk
of a cloned transgene cow at levels of up to 5 g/l
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Hormone transgenic livestock.Transgene then germline of animals.mans.
Benefit farm animals and humans.
However restricted inefficiencies methodology.
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Implantation __________________________________________________________________________________
Livestock costly.
Incorporated foreig DNA sperm mediated gene transfer SMGT.
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Phytate phosphorus pigs and chicken.
produce enzyme phytase in thesalivary glan secreted in the saliva.
100,000 untis/kg.___________________________________________________________________________________
Phosphorus in feces enzyme stable and active degrade in pancreatic. Protease.
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Transgenic dogs Alzheimers disease breakthrough. GMO animal models for alzheimers disease
Brain disorder , pharmaceutical. Industry.
Expressinng plant genes.Levels of unsaturated fatty acids > 100 fold higher
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Preadipocites derived from adipose tissue at transgenic fish cultured in vitro than intact fat tissue from
.
__________________________________________________________________________________Transgenic sheep anf goat .
Milk produced a human protein alpha antitrypain,
Treat aquatic fibrosis , 1st clone dolly the sheep.
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Human factor IX transgenic sheep produced by transfer of nuclei.
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Improve wool pronucleus microinjection the high-sulfur keratin
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Golden egg a new way to make drug.
Eggs and human protein gene alter chicken.
Antibody used for passive immunization for diagnostic purposes like blood group , disease testingTherapeutic intervention.
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Monoclonal antibody mab
Fusion of B-lymphocytes immune cells of a mouse.
Myeloma cancer cell.
Hybridoma immortal cancer cell and specific antibody.
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Mab
Purification process.
es s ma recogn ze mo ecu e.
Pregnancy , fertility test.Safe
Broad range of potential targets.
High selectivty.
High affinity.
Ease of preparation._______________________________________________________________________________
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Therapeutic
High therapeutic dose.
Low immunogenicity.Poor penetration in solid tumors.
Potential cross reactivity and other tumors.
High production cost.
Potential viral safety problems.
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All mab Abs were derived from rodent cells
Rodent-derived mab suffered from short half life and poor recognition of the rodent IgG-C region
Human effectors function.
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enera on o rue uman ma s ns a e an on y m e num er o arge s.
-Chimerization resulting mab 60-70%-Humanication 90-95%.
-Use phage display fully mab.
-Transgenics fully human mabs.
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The rDNA technology
Insulin from E.coliMore complex and larger gene products > insulins
Can not post translational.
Glycosylation.
Folding.
TrimmimgBiological function.
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Protein
Bioactivity erythropoietin , tissue plasminogen .activation.
Development of posttranslational processing.
Development of cell line.
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E.Coli for animal cells.
Biopharmaceutical was produced in E.coli. 86%50-60% produced in animal cells CHO Hamster.
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-The development rDNA heterogenous gene expression.
-The disadvantage is the high production costs and production in cultivated bacteria , yeast cells.
- Research program have developed robust and highproducing animals host cell lines._______________________________________________________________________________
Animal cell proteins.
-Therapeutic biopharmaceuticals using animal cell technology.
-proteins used for the treatment of cardiovascular disease.
tPA ,tissue , plasminogen activation.
Cystic fibrosis Dnase , erythropoietin EPO.Viral infection , interferon and interleukin.
,multiple sclerosis , interferon-beta ,
Human growth ,GH.
.
Culturing of animal cells farms also an important of gene therapy.Delivery gene to patient.
Result long lasting treatment , recombinant viruses.
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Drug discovery.
The evolution of drug discovery and development process has been revolutionized by high-throughput
Technology.
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-Chemistry.
Gemomics-based target identification.
Animals and human cells.Cell-based assays and models.
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Large scale cultivation for industrial purposes is performed in bioreactors large stainless steel tanks and
Gentle agitation and aeration to supply o2.
Cells , multiple,release protein into medium.
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Human protein.
Product sufficient amount.
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a er or e env ronmen .
__________________________________________________________________________________The vector used
Retroviral vectors.
Adenoviral vextors.
Naked , plasmid DNA.
Lipofection.
Pox viruses vector.
Vaccination viruses vectors.
Herpes simplex virus.
AAV adevo-associated viruses.
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Preclinical studies to evaluate, number of gene therapy,gene therapy program for neuologic illnesses,
Autoimmune diseases, allergies, regeneration of tissues.
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