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Biotechnology Genetic Engineering The use of microorganisms or biological substances to perform specific industrial or manufacturing processes Recombinant DNA Genes from different species are combined and placed into host cells • Example: • Green Fluorescent Protein (GFP) • Found in jellyfish, fireflies Making Recombinant DNA 1. Isolate gene of interest • “Cut” the GFP gene from a jellyfish’s DNA using a restriction enzyme • Each restriction site codes for a SPECIFIC restriction enzyme 2. Insert gene of interest into a plasmid • (GFP) gene + vector= recombinant plasmid • Vector is a small, circular piece of DNA • Plasmid transports target (GFP) gene to host cell 3. Transfer plasmid into cells of host organism GFP Plasmid gets inserted into EMBRYONIC cat cells Cat’s Chromosomes within nucleus Plasmid 4. Transformation of host organism’s cells •Plasmid will eventually be incorporated into host organism’s DNA •In our example, cat will now be able to produce GFP • AKA the cat will now GLOW!!! Mr. Green Genes, a transgenic cat Green fluorescence protein, or GFP, was inserted into his genome using recombinant DNA technology. GFP fluoresces (glows) when exposed to a blacklight Other uses for recombinant DNA: 1. Agriculture: -Crops: Disease protection, drought & frost resistance, increased growth -Livestock: Increased muscle mass, hormone acceleration, drugs in milk 2. Pharmaceuticals: HGH, Insulin: insertion of human gene into bacteria for cheap proteins. Instead of putting plasmid into eukaryotic cells, you can put it in bacteria, which will undergo transformation, and produce a bunch of the protein for human use INSULIN PRODUCTION! Other applications of biotechnology… Genetically modified food These fish are the same age, but the one on the bottom is growing much slower. The fish on the top has been genetically modified to grow faster. These tomatoes were grown in the same field. The tomatoes on the left have been eaten by pests while the tomatoes on the left are untouched because they were genetically modified to be pest resistant. Pros Genetically Modified (GM) crops can be made resistant to viruses, fungi and bacterial growth. -grow faster. -pest-resistant -tolerate extreme weather conditions -engineered with added vitamins and minerals Cons What are the long term health effects? Can GM foods cause new allergies? What are the effects on the environment? What’s next…genetically modified animals? How can you tell your food is genetically modified? A warning label is currently not required To know, look for common GM ingredients Soy lecithin Fructose Dextrose Grapeseed oil Cottonseed oil Most corn syrups or corn starches Some foods are labeled GMO Free More common than you think…. DNA can be used in Crime Scene Investigation DNA Fingerprinting • Each person has their own unique DNA “fingerprint” except for identical twins • Used for forensics, paternity testing, and identification of remains • Small quantities of blood, semen, or body tissue can be tested for DNA base sequences DNA Fingerprints are made by Gel Electrophoresis Gel Electrophoresis is a technique used to separate molecules based on size The size of the fragments are unique to each person so it creates a DNA fingerprint How does gel electrophoresis work? 1. Restriction enzymes cut DNA 2. Pieces of DNA are run through a gel 3. The pieces of DNA are separated by weight and make a banding pattern that is unique to an individual (fingerprint) 4. Bands that align with each other indicate a match Gel Electrophoresis A case for Maury • Shown at the right are DNA "fingerprints" from a new mother (M), her recently born child (C), and two possible fathers (F1, Ross and F2, Rick). • What do you think is Ross (F1) or Rick (F2) the father of the child? M C F1 F2 A crime scene case • Shown to the right are DNA "fingerprints" taken from seven (7) suspects (the numbered columns). • The bloodstain in the middle is from the crime scene, ostensibly from the killer. Which suspect is guilty? How many genes do humans have? What do they code for? Human Genome Project 1990-2003 Goals: Identify ALL genes in human DNA Determine sequence of the base pairs We thought there were BILLIONS of bases in the human genome Store all information in database Use information for disease/drug research, to compare human genome to other organisms Obama, State of the Union 2013: “For every $1 we spent mapping the human genome, we got $104 back” With the HGP, we have discovered which genes are affected when you have the above diseases What did we learn? We only have about 30,000 genes A huge portion of our genome is non-coding, meaning it does not code for proteins In order to make huge advances in diseases like cancer, the environment has to be taken into account Gave rise to fields such as epigenetics Is it possible to fix a genetic mutation using biotechnology?? …Yes! It’s called gene therapy! Gene Therapy • Replacement of defective genes with healthy genes • Prevents or treats disease • Examples: cystic fibrosis, hemophilia, sickle-cell anemia, immune-deficiencies Pin Egg cell Cloning Needle Cloning is the production of genetically identical organisms from single cells First gene cloned mammal= Dolly the sheep How it works… Egg cell is enucleated (nucleus removed) and replaced with a nucleus taken from an adult somatic cell The egg is then placed in the reproductive system of a surrogate mother, where it develops normally Surrogate gives birth to offspring that is http://learn.genetics.utah.edu /content/cloning/whatisclonin identical to nuclear donor g/ Animals that have been cloned • • • • • • • Fish Cats Cattle Deer Dog Ferret Fruit Flies • Goat • Horse •Mice •Mule •Pig •Rabbit •Rat •Rhesus Monkey •Sheep •Water buffalo •Wolf Pets cloned These dogs, Mira and Missytoo, were both cloned from a dog that died in 2002. They are owned by Lou Hawthorne of California. Mr. Hawthorne owns BioArts, a company with offices in California and laboratories in South Korea. The company offers pet cloning for around $150,000. Current debate December 28th, 2007 - FDA said cloned animals are safe to produce and eat Waiting for consumer approval - has not happened yet United States bans human cloning of all types including organs. Stem Cells Is it possible to grow a leg or an arm from just one cell? Stem Cells Stem cells are cells that are undifferentiated They can become any cell, in any part of the body, and can divide without limit to replenish other cells as long as the person is alive Found in all multi-cellular organisms 2 types of stem cells Embryonic: taken from a fertilized egg, about 4-5 days old. They can become any of your 200 cell types. Somatic: Found in bone marrow and the umbilical cord. Not as effective as embryonic stem cells and are limited to what they can become. What are the ultimate goals of stem cell research. Uses of stem cells MANY !!!!!!!!!!!!!! Possible cure for many genetic diseases Alzheimer's Cancers Parkinson's Limb, organ re-growth Spinal cord injuries Hamburgers?! The controversy Embryonic Requires destruction of a human embryo (zygote) Recently single cell biopsies have been done that can allow cell creation without embryo destruction!