3 Biotechnology Fermentation Animal cloning Genetically modified foods and the American-European opinion divide.Animal cloningGene cloning for pharmaceutical productionDNA fingerprintingThe promise and perhaps perils of embryonic stem cells
4 1. FermentationA technique that requires bioreactors to allow a biological process under controlled (optimum) conditions producing a useful substance in large amountAlcoholic drinksProduction of wine and beerRecently mammalian cells
5 Can grow trillions of new bacteria One cell with the recombinant plasmidCan grow trillions of new bacteriaA fermentor used to grow recombinant bacteria.
6 2. Biotransformationchemical modifications made by an organism on a chemical compound.Microbial BiotransformationBiodegradationBioremediationMineralizationBiosurfactant
7 Microbial biotransformation Biotransformation of various pollutants to clean up contaminated environmentsThe methods harness the naturally occurring, microbial catabolic diversity to degrade, transform or accumulate a huge range of compounds including hydrocarbons (e.g.oil,PCBs (polychlorinated biphenyl),PAHs (polyaromatic hydrocarbons),pharmaceutical substances, andmetals.Biological processes play a major role in the removal of contaminants and pollutants from the environment .
8 BiodegradationProcess by which organic substances are broken down by the enzymes produced by living organisms.The term is often used in relation to ecology, waste management and environmental remediation, bioremediationOrganic material can be degraded aerobically, or anaerobically.
9 BioremediationAny process that uses microorganisms, fungi, green plants or their enzymes to return the natural environment altered by contaminants to its original condition.Bioremediation may be employed to attack specific soil contaminants, such as degradation of chlorinated hydrocarbons by bacteria e.g.cleanup of oil spills by the addition of nitrate and/or sulfate fertilizers to facilitate the decomposition of crude oil by indigenous or exogenous bacteria.
10 MineralizationThe process where a substance is converted from an organic substance to an inorganic substance.modification ends in mineral compounds like (NH+3 or H2O),This may also be a normal biological process which takes place during the life of an organism such as the formation of bone tissue or egg shells, largely with calciumThis term may also be used to indicate the digestion process in which bacteria utilize the organic part of the matter, leaving behind the minerals
11 Biosurfactants Surface-active substances synthesized by living cells. They have the properties of reducing surface tension, stabilizing emulsions, promoting foaming and are generally non-toxic and biodegradable.Biosurfactant producing microorganisms may play an important role in the accelerated bioremediation of hydrocarbon contaminated sites.These compounds can also be used in enhanced oil recovery and may be considered for other potential applications in environmental protection
12 3. Cell fusionInvolves combining two cells to make a single cell that contains all the genetic material of the original cells.Used to create new plants by fusing cells from species that do not hybridize naturallyCrossbreeding and then generation of new plant from the fused cell e.g. POMATO (fusion of tomato and potato)Used to generate monoclonal antibodies(Protective proteins produced by a clone of a single cells)
13 How cells can fuse? Using Viruses Using chemicals (polymers) Weaken the cell wall of the cells and cause them to bind togetherUsing EnzymesPlant cells can be fused only after removing the cell walls completely by enzymatic reactions
15 4. LiposomesMicroscopic spherical particles that are formed when lipids form a suspension in water.Lipid molecules are arranged in a manner so as to enclose a tiny space in the centre to carry a drug or any other materialUsed as vehicles for delivering certain drugs to the target tissues of the bodyDrugs are encapsulated in liposomesprotect against digestive enzymes in the stomach
16 The spontaneous closure of a phospholipid bilayer to form a sealed compartment. Closed structure is stable because it avoids the exposure of the hydrophobic hydrocarbon tails to water (energetically unfavorable.
17 PHOSPHOLIPID BILAYERS A bilayer of phospholipids forms a sphere in which water is trapped inside. The hydrophilic phosphate regions interact with the water inside and outside of the sphere.The fatty acids of the phospholipids interact and form a hydrophobic center of the bilayer.
18 Liposomes (A) An electron micrograph of unfixed, unstained phospholipid vesicles. (B) Schematic representation of lipososmes
19 5. Cell and tissue culture Growth of living cells or organism outside the body in a suitable culture medium which provides nutrients to the growing cells.Individual cells grow and divide in s sterile medium.Extensively used in Labs e.g.Cancer ResearchPlant breedingKaryotyping (chromosomal analysis)Organized profile of a person’s chromosomes
20 6. Genetic EngineeringThe transfer of genes (segment of DNA) from one species to another.Requires recombinant DNA technologyGene transfer techniquesUsed to improve breeds of economically important plants, cow, gorses, dogs, etc.
21 Biogenetic Techniques Gene Splicing Isolation of DNA molecules,Cutting of DNARestriction AnalysisRejoining of DNA molecules from different sourcesFormation of recombinantsTransformation
22 Transformation?Introduction of foreign DNA into the cells (Prokaryotic cells, bacteria) using non-viral methodsConversion of normal cell into cancerous cell
23 Transfection?Transfection is the process of introducing nucleic acids into cells (eukaryotic cells such as fungi, algae and plants) by non-viral methods.
24 Transduction?Naturally, process by which DNA is transferred from one bacterium to another by a virus.In biotechnology, it refers to the process whereby foreign DNA is introduced into another cell via a viral vector.
25 Gene TherapyInsertion of normal gene from one organism into the patient’s cells (where gene is defective)Post-transcriptional gene silencing using small pieces of RNA (RNAi)
26 DNA Fingerprinting (DNA typing, DNA testing etc.) To identify various components of DNA, unique for an individualA portion of the DNA of an organism uniquely distinguishes that organism from anotherTechnique is based on mini-satellites, repeated several times in the genome
27 DNA FootprintingDetects DNA-protein interaction using the fact that a protein bound to DNA will protect that DNA from enzymatic cleavage.Using this technique, it is possible to locate a protein binding site on a particular DNA molecule.
28 Gene Mapping Mapping of genes to specific location on chromosome Assigning DNA fragments to chromosomes
29 In vitro; within the glass, outside the living system Performing a given procedure in a controlled environment outside of a living organismIn vivo; within the livingExperimentation using a whole, living organism as opposed to a partial or dead organismIn situ; in the place (usually means something intermediate between in vivo and in vitro).Examining a cell within a whole organ intactEx vivo; out of the livingexperimentation or measurements done in or on living tissue in an artificial environment outside the organism with the minimum alteration of the natural conditions
30 CloningA technique used for production of organism from a single individual, which are identical in genetic constitution.A clone is an organism that is derived from a single parent through non-sexual methods.PlantsMicroorganismsNot in mammalsCloning of mammals (Nuclear Transfer)as practiced in Dolly at Roslin Institute of EdinburghDolly was unable to breed and produce healthy offsprings
31 Embryo Transfer To improve dairy cattle Artificial insemination Surgical removal of a fertilized eggTransfer into a recipient female
32 Cloning Technologies Recombinant DNA technology DNA cloning Molecular cloningGene cloningReproductive cloningTherapeutic cloningEmbryo cloning
34 The Biotechnology of Reproductive Cloning Even under the best of circumstances, the current technology of cloning is very inefficient.Cloning provides the most direct demonstration that all cells of an individual share a common genetic blueprint.
35 Why Clone Animals? To answer questions of basic biology Five genetically identical cloned pigs.To answer questions of basic biologyFor pharmaceutical production.For herd improvement.To satisfy our desires (e.g. pet cloning).
36 Is Animal Cloning Ethical? The first cloned horse and her surrogate mother/genetic twin.As with many important questions, the answer is beyond the scope of science.
37 Saved by Cloning?Some are firm believers while many view these approaches to be more of a stunt.Note the use of a closely related species, a domestic goat, as egg donor and surrogate mother.
38 Carbon Copy– the First Cloned Pet (Science (2002) 295:1443)Significantly, Carbon Copy is not a phenotypic carbon copy of the animal she was cloned from.
39 DNA CloningTransfer of a DNA fragment of interest from one organism to a self-replicating genetic element such as a bacterial plasmidPlasmidsSelf-replicating extra-chromosomal circular DNA molecules, distinct from normal bacterial genome
40 DNA Cloning - UsesGene therapyGenetic engineering of organismsGenome sequencing
41 Reproductive CloningA technology used to generate an animal that has same nuclear DNA as another currently or previously existing animalDollyHow Is Reproductive Cloning Done?Somatic cell nuclear transfer (SCNT)
42 Therapeutic Cloning Production of human embryos for use in research GoalTo harvest stem cells that can be used to study human development and to treat disease
43 Genetically Modiﬁed Cloned Offspring Biotechnological applicationsProduction of pharmaceuticalsXenotransplantationStudy and eradication of human diseaseImprovement of livestock
44 Production of Pharmaceuticals Insulin for diabetesInterferon for viral infectionsTissue plasminogen activator (which dissolves blood clots)
45 XenotransplantationTo develop animals whose organs will not cause an immunological response and destroy transplanted tissue when transferred to humansPig organsHearts, lungs, kidneys, liverNeural tissue for Parkinson'sIslets cells for diabetes patients
46 Study Human Disease Sheep model to investigate human cystic ﬁbrosis Cloned sheep used for drug testing and to evaluate new therapies
47 Improvement Of Livestock Cloning animal with excellent traitsProduction of a large number of clones from high quality animalsAllow overall genetic improvement of herdRepopulate endangered animals
48 Animal Cloning Sheep, cattle, goats, pigs, and mice Cloning efforts in rabbits, rats, cats, dogs, and horses are ongoing
49 Risks Of Cloning Reproductive cloning expensive and highly inefficient > 90% of cloning attempts fail to produce viable offspring
50 The Next Step? Highly unlikely Attempts at human cloning are viewed very unfavorably in the scientific community