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Biotechnology AP Biology 2007-2008 Biotechnology today Genetic Engineering Electrophoresis Recombinant Technology Polymerase Chain Reaction Transgenics/GMO’s Cloning AP Biology Recombinant Technology rDNA is DNA with 2 or more sources Bacteria are used to copy DNA and make desired proteins Identify gene of interest and cut it out insert new gene into a plasmid insert plasmid into bacteria Vector – plasmid carrying new gene into host AP Biology bacteria can express new gene or make copies of inserted DNA AP Biology Restriction Enzymes – Cut the DNA! a.k.a.restriction endonucleases evolved in bacteria to cut up foreign DNA from viruses Molecular scissors that cut between specific bases “Sticky Ends” are where the enzyme cut allow for joining with another piece of DNA cut with the same enzyme AP Biology Recombinant tech. Transformation insert recombinant plasmid into bacteria grow recombinant bacteria in agar cultures bacteria make lots of copies of plasmid “cloning” the plasmid production of many copies of inserted gene production of “new” protein transformed phenotype DNA RNA protein trait AP Biology Recombinant Plasmid Plasmids used to insert new genes into bacteria cut DNA gene we want like what? …insulin …HGH …lactase cut plasmid DNA Cut DNA? DNA scissors? ligase recombinant APplasmid Biology insert “gene we want” into plasmid... “glue” together Grow bacteria…make more gene from other organism recombinant plasmid + vector plasmid grow bacteria harvest (purify) protein AP Biology transformed bacteria Transgenics – organisms with foreign DNA Genetically modified organisms (GMO) enabling plants to produce new proteins Protect crops from insects: BT corn corn produces a bacterial toxin that kills corn borer (caterpillar pest of corn) Extend growing season: fishberries strawberries with an anti-freezing gene from flounder Improve quality of food: golden rice rice producing vitamin A improves nutritional value AP Biology Green with envy?? Jelly fish “GFP” AP Biology Transformed vertebrates Many uses of restriction enzymes… Now that we can cut DNA with restriction enzymes… we can cut up DNA from different people… or different organisms… and compare it why? forensics medical diagnostics paternity evolutionary relationships and more… AP Biology Copy DNA without plasmids? PCR! Polymerase Chain Reaction method for making many, many copies of a specific segment of DNA ~only need 1 cell of DNA to start It’s a DNA copy machine! AP Biology PCR process What do you need to do? in tube: DNA, DNA polymerase enzyme, primer, nucleotides denature DNA: heat (90°C) DNA to separate strands anneal DNA: cool to hybridize with primers & build DNA (extension) What does 90°C do to our DNA polymerase? AP Biology Comparing cut up DNA How do we compare DNA fragments? separate fragments by size How do we separate DNA fragments? run it through a gelatin agarose made from algae gel electrophoresis DNA jello?? Can’t we just add those little marshmallows? AP Biology Gel electrophoresis DNA moves in an electrical field… DNA is negatively charged size of DNA fragment affects how far it travels small pieces travel farther large pieces travel slower & lag behind DNA – AP Biology “swimming through Jello” + Gel Electrophoresis DNA & restriction enzyme longer fragments wells power source gel shorter fragments AP Biology + completed gel Uses: Evolutionary relationships Comparing DNA samples from different organisms to measure evolutionary relationships turtle snake rat squirrel fruitfly – DNA + AP Biology 1 2 3 4 5 1 2 3 4 5 Uses: Medical diagnostic Comparing normal allele to disease allele chromosome with normal allele 1 chromosome with disease-causing allele 2 – DNA Example: test for Huntington’s disease + AP Biology Uses: Forensics Comparing DNA sample from crime scene with suspects & victim suspects S1 S2 S3 crime scene V sample – DNA AP Biology + Electrophoresis use in forensics Evidence from murder trial Do you think the suspect is guilty? blood sample 1 from crime scene blood sample 2 from crime scene blood sample 3 from crime scene “standard” blood sample from suspect OJ Simpson blood sample from victim 1 N Brown blood sample from victim 2 R Goldman AP Biology “standard” Uses: Paternity Who’s the father? Mom F1 – DNA AP Biology + F2 child Clone A genetically identical organism or a group of cells derived from a single cell AP Biology Cloning Process The nucleus from a somatic (body) cell is harvested An egg is harvested and the nucleus of the egg is removed The nucleus from the somatic cell is then transferred into the egg An electric shock causes the cell to replicate The embryo is now implanted into the uterus to grow into a new individual AP Biology Big Ideas of Biotechnology Genes from virtually any organism can be cloned using plasmid transformation or using PCR. Electrophoresis and restriction enzymes are widely used in DNA analysis. GMO’s and cloned animals and plants can be given beneficial characteristics or make needed products such as pharmaceuticals. AP Biology