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28 Cards in this Set
- Front
- Back
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Biotechnology
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Application of biological systems(microorganisms) to obtain a product(food, antibiotics)
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Recombinant DNA technology
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procedures by which a fragment of DNA gene of one organism is incorporated into a genome
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Goals of Genetic Engineering
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-Eliminate undesirable plenotypic traits in humans, animals, and microbes
-create organisms that synthesize products humans need(insulin) |
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Tools/Techniques of Genetic Engineering
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-Restriction enzymes-Major Tool
-Analysis of DNA-gel electrophoresis -nucleic acid hybridization -DNA sequencing -Polymerase chain reaction-PCR |
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Restriction Enzymes
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-DNA cutting enzymes
-they recognize and cut fragments of DNA -Leave single stranded sticky ends of DNA -Same restriction enzyme =same sticky end -Cut on specific Palindromic Sequence( identical both ways) |
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DNA Gel Electrophoresis
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-separation of DNA fragments based on size
-in agarose gel DNA fragments are subjected to electrical current -Molecules with negative charge move to positive pole -smaller fragments move faster |
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Nucleic Acid hybridization
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fragment of single stranded nucleic acid can unite with fragment of complimentary seqeunce
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Hybridization with Probe
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-method used to detect specific nucleotide sequence in unknown sample by using gene probe
-probes carry a radioactive label |
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DNA sequencing
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-process in which exact sequence of nucleotides in a DNA segment is determined
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Polymerase Chain Reaction- PCR
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-small amount of specific DNA fragment can be amplified in vitro with PCR machine(thermal agar)
-Target DNA serves as template -supply 4 nucleotides -DNA polymerase -primers-short fragments of DNA complimentary to target DNA |
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PCR Cycle -3steps
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-Denaturation: at 94degrees C separates DNA strands
-Priming: 50-65 degrees primer attached to complimentary strand of DNA -Extension: 72 degrees DNA polymerase extends the molecule by adding nucleotide. Millions of DNA copies |
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Recombinant DNA technology
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-an elected gene is removed from genetic donor
-gene is incorporated into a vector(plasmid of virus) -vector inserted into a cloning host (bacteria, yeast) -incorporated in segments in which foreign DNA can be( plasmids,viruses) |
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Vector Characteristics
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-self-replicating
-circular shape -proper size to accept foreign DNA -must have promoter -gene for antibacterial resistance |
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Transformation
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-plasmid from surrounding enviroment is taken up by cell
-cells have too be made competent by soaking in calcium chloride |
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What does vectors(plasmids) contain?
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gene for ampicilln resistance
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What can only cells that have been transformed can do?
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grow on medium containing ampicillin
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Synthetic DNA
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-DNA synthesis machine
-short strands of DNA can be synthesized (120 nucleotides) -must know segment sequence wished to be synthesized |
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Applications of recombinant DNA technology
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-pharmaceutical/theraputic
-protein synthesis(vaccines, DNA vaccines, gene screening) DNA finger prints(gene therapy) |
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Example of recombinant technology
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-hormone insulin for diabetics
-gene was cloned into e.coli -obtained from pancreas -1st success of recombinant |
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Somatostatin
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-treatment of giantism
-before 500,00 sheep brian needed for 5mg of somastatin -today 81 needed for equilvalent |
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Subunit Vaccines
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-protein portion of virus is cloned
-Hep B vac -Advantage: no chance of becoming infected |
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DNA vaccines
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-Single gene from artifically copied and multiplied pathogen
-gene injected into muscle. and muscle take gene as one of own -immune system recognize as foreign gene and produce antibodies |
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Agricultural Applications
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-transgenic organisms altered by addition of genes from other organisms
-improving crops |
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Creating transgenic plants
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-agrobacterium tumefaciens-plant parasite that incorporates its DNA by Ti plasmid
-Ti plasmid can be engineered to contain new gene |
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Resistance to Herbicides
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-Glyphosate(round up)kills all plants
-gene for resistance to glyphosate in crop plants -kill weeds not crops |
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Transgenic Animals
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-many human genes have better expressions in animals than in bacteria
-the protein can be collected in milk or semen -foreign gene into embryo by virus or injection |
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Gene Therapy
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-missing or defective gene can be replaced with normal copies
-treatment for cystic fibriosis, sicle cell -can be researched to treat cancer |
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Genetic Screening
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-genetic diseases detected by genetic engineering
-technique:southern blotting -inherited forms of breast cancer |
