PLB 115 1st Edition Biotechnology Lecture 14Previous Notes’ OutlineI. DNA StructureII. DNA FunctionsIII. DNA ReplicationIV. Base Pairing ProcessV. DNA CodeVI. DNA VS RNAVII. Protein SynthesisVIII. TranscriptionIX. TranslationX. Ribosomes & TranslationXI. Translation InitiationXII. Translation ElongationXIII. Translation TerminationXIV. Universal Genetic CodeXV. Control of Protein SynthesisXVI. Control of Protein QualityXVII. Alternative SplicingXVIII. Mutations & Protein SynthesisXIX. Point MutationsXX. Sickle Cell AnemiaXXI. Insertions & DeletionsXXII. Mutations Caused by VirusesCurrent Notes’ OutlineI. BiotechnologyII. DNA FingerprintingIII. DNA Fingerprinting TechniqueIV. Polymerase chain reactionV. ElectrophoresisVI. Variable Number Tandem Repeats (VNTR)VII. Genetic Modification of OrganismsVIII. How to Clone a GeneIX. Insulin from BacteriaX. Purpose of Genetically Modified OrganismsThese notes represent a detailed interpretation of the professor’s lecture. GradeBuddy is best used as a supplement to your own notes, not as a substitute.XI. Cloning OrganismsXII. Stem CellsXIII. Personalized Stem Cell LinesXIV. EthicsBiotechnologyBiotechnology: Collection of laboratory techniques that involve direct manipulation of an organism’s DNA.Examples of its use: - Creation of cheaper & more effective drugs - Correction of genetic mutations- Creation of cells that can clean-up environmental messes - Creation of an increase in agricultural productivityBiotechnology involves the manipulation of DNA in order to change phenotype- Organisms that have similar phenotypes have similar DNA sequenceso DNA can be compared in 2 ways: DNA fingerprinting & DNA sequencingDNA FingerprintingUniquely identifies individuals on the basis of DNA fragment lengths- Fragments are generated by restriction enzymes that cut DNA at specific siteso Each individual’s DNA is different enough that these enzymes will generate different lengths of fragments in two different individuals Can identify if two samples of DNA came from the same person- For crime scenes, paternity casesDNA Fingerprinting Technique DNA is obtained from: - Blood at a crime scene - Hair from crime suspect- Polymerase chain reaction is used to make many copies of a VNTR (variable number tandem repeat) region of DNA- Restriction enzymes are used to cut the VNTRs into fragments- Fragments are separated by electrophoresis- Patterns of the two DNA samples are compared-Polymerase chain reactionDNA is replicated in a test tube - Uses primers that dictate the region of DNA to be copied - Makes many copies of a particular segment of DNAo Creates enough material for scientists to perform DNA fingerprinting-ElectrophoresisA way to separate DNA fragments based on their length- DNA sample is loaded into a gel matrix and an electrical current is applied- Smaller fragments travel through the gel faster- Creates a banding pattern of fragmentsVariable Number Tandem Repeats (VNTR)Regions of DNA that are repetitive sequences- Each person has a slightly different number of repeatso Regions are cut with restriction enzymes & each person has a different set of fragmentsGenetic Modification of OrganismsGene cloning: Cloning genes & moving them from one organism to anotherCreating recombinant DNA: DNA sequences altered (mutated) to generate a desired change- DNA is transferred & new host cell begins to make the new DNA & produce new proteinso Usually involves bacteria or viruses that will make large amounts of the protein ofinterestHow to Clone a Gene- Cut the gene of interest out of the chromosome using restriction enzymes- Splice the fragment containing the gene into a carrier molecule (bacterial plasmid)- Insert the plasmid with the fragment into bacterial cells- Each time bacterial cells divide, many copies of the gene will be madeInsulin from BacteriaPurpose of Genetically Modified Organisms- Make human insulin - Generate “Golden rice” - Make interferon - Make human growth hormone- For bioremediation (the use of living organisms to remove toxins from the environment)- Generate crops that supply developing nations with nutrients not normally found in their native plants- Generate crops that can manufacture medicines to treat disease- Generate crops that are resistant to herbicides or that make their own insecticidesGene Therapy: Manipulating genes in order to cure or treat a genetic diseaseGene therapies must be specifically designed for each situation- If the mutant gene is not functional, then a functional gene must be inserted- If the mutant gene is overactive, then it must be deleted or alteredo Usually involves mutating the part of the gene that controls its activationCloning OrganismsSomatic cell nuclear transfer- A nucleus must be removed from a cell of the organism that is to be cloned- The nucleus is then placed into an enucleated egg- The egg is stimulated to divide and is artificially implanted into a host motherCloning has advanced our understanding of developmental processes like…- Determination is the process a cell goes through to select which genes it will express- Differentiation is the process of expressing those genes and becoming a specific cell type**Cloning of Dolly (sheep) showed that determination can be reversed & a fully differentiated cell can be used to clone an organism**Stem CellsCells that haven’t completed the determination & differentiation process, so they have the potential to develop into many/any different cell types - Could be used to replace damaged tissues and organs in humans& cure diseaseso Parkinson’so DiabetesEmbryonic stem cells come from embryos & have great potential ** However, embryos must be destroyed (die) in order to obtain them and work with them**Adult stem cells have partially completed the process of determination- Have less potential; they are already specific tissue types (blood, bone, etc.)Personalized Stem Cell LinesCombining somatic cell nuclear transfer & stem cells may allow scientists to create personalized stem cell lines- The individual’s nucleus would be transferred to an egg- The egg would be allowed to divide to the point where it produced embryonic stem cells- Could potentially be used to grow tissues or organsEthics Concerning Biotechnology- QuestionsWill the technology be used safely? Who will benefit?Who will suffer? Should the technology be used to make a profit?Just because we can, should