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Unit 6 BIOTECHNOLOGY Maury http://www.youtube.com/watch?v=ZD78- CCPhAQ What did they do to tell who the father was? How do they do these tests? How do they know for SURE? What is biotechnology? Bio = Life Technology = machinery, equipment, or the application of knowledge Technology that relates to the field of biology or living things Manipulating living organisms using various types of technology to produce more useful qualities using better genes and traits Allows the best or desired traits to be expressed. Research in Biotechnology Help humans create crops that can be frost resistant 2. Use transgenic organisms to help medical researchers model human physiology for testing 3. Help industries to create bacteria to break down pollutants into harmless products 4. Pharmaceutical companies use recombinant DNA to cheaply produce human hormones (insulin) and other proteins 5. Help solve crimes and determine familial relationships 1. Gel Electrophoresis A method used in biochemistry and molecular biology to separate DNA or RNA molecules by size to compare DNA data. Also called a “DNA fingerprint” Unique to each person What can this be used for? Paternity Tests Who is the father? Familial Relationships Who is the mother? Who are the siblings, cousins, grandparents, etc.? Evolutionary Relationships How closely related are two organisms (humans & apes)? Crime Scene Investigations Who committed the crime? Basic Steps.. 1. Extract DNA sample 2. Cut DNA into smaller pieces using Restriction Enzymes 3. Put the smaller DNA fragments into the Gel Tray 4. Use an electric charge to separate the pieces of DNA 5. Analyze the DNA fragments Goal of Gel Electrophoresis 1. The process of Gel Electrophoresis works to separate and analyze DNA fragments according to their size & shape. 2. In order to cut the DNA into fragments, restriction enzymes are used. These are specific enzymes (proteins) that recognize specific sequences of DNA and then cut it in half. 3. Samples of DNA fragments are loaded onto a GEL ELECTROPHORESIS. 4. DNA fragments move in the Gel due to opposite charges (DNA is negatively charged) The SMALL strands move further down the Gel than the LARGER strands of DNA. Since they are larger and heavier, the LARGER strands do not move as far down the Gel 5. The groups of DNA fragments show up as small lines in the Gel 6. Now a DNA FINGERPRINT is created, which can be analyzed. Let’s Take a look… http://www.youtube.com/watch?v=PSwlCk_Z 02c&feature=related Check Point 1. What are the steps used in Gel electrophoresis? 2. Where would you find the shortest stand of DNA? 3. What causes the fragments on DNA to move? DNA fingerprinting applications Sample Problems Genetic Engineering – Transgenic Organisms What is a transgenic organism? Let’s break down the word… Trans = across Genic = genes Organism = Living thing Transgenic Organisms are: It is an organism that has had genes inserted (or moved into) from a different organism Transgenic organisms are made from combining different DNA Genetically Modified Food (GMO) Genetically Modified Food (GMO) What is Genetically Modified Food (GMO)? It is a crop or food that has been intentionally modified, altered or changed. What is Selective Breeding… A type of genetic engineering Choosing organisms (plants, animals) with the best traits (genes) and breeding them together to produce better (bigger, stronger) offspring Can you thinks of a real life example of selective breeding? How did THIS happen?! Glow-in-the-dark PIGS?! A desired gene from one organism (the glow gene from a fire fly) was inserted into the DNA of a pig Through MITOSIS the pig’s cells began replicating the glow-in-the-dark gene (the desired gene) Let’s take a closer look at this… What is Recombinant DNA When DNA from two different organisms come together, the DNA becomes RECOMBINANT DNA What does recombinant (recombine) mean? • The organism that receives the recombinant DNA will use it as if it were its own DNA. Also know as… Transgenic Organisms How do scientists create DNA? Its all about using BACTERIA! This is because bacteria have DNA rings called PLASMIDS. Steps for Creating a Transgenic Organism 1. Remove the plasmid 2. Cut the bacterial DNA using restriction enzymes, 3. 4. 5. 6. leaving an open “sticky end.” Remove and cut a desired gene from a different organisms (foreign DNA), also leaving an open “sticky end.” Then glue a piece of foreign (human, plant, animal) DNA back into the plasmid using the enzyme Ligase and sticking the “sticky ends” together. Insert the plasmid back into the Bacteria Now, the bacteria will reproduce the new gene (in the plasmid DNA) to make larger quantities of the desired gene or trait. All bacteria are… IDENTICAL REMEMBER: bacteria divide using asexual reproduction! SO… Every new bacteria will be making exact copies of the desired gene! Applications of Recombinant DNA Bacteria used to produce hormones, antibiotics and Insulin Plants made to be resistant to herbicides and Pesticides Animals Cloning Replacement of genes which cause disorders (aka Gene Therapy) Such as Cystic Fibrosis Severe Combined Immunodeficiency Check Point 1 1. What is a Transgenic Organism? 2. How are transgenic organisms made? 3. What is a GMO? 4. What is a real-life example of a GMO? 5. What is selective breeding? 6. What is a real-life example of selective breeding? Human Genome Project A collaborative effort by scientists around the world to map the human genome Genome = all of an organism’s hereditary information Completed in 2003 Can be used to determine if people carry genes for certain traits or diseases Can be used to determine the locations of genes on chromosomes Problems with the Human Genome Project: What do we do with the information? Designer babies? Abortion of defective babies? Sterilize carriers? Will insurance companies be able to deny you coverage because of your genes? Can pharmaceutical companies “copyright” a gene sequence? If so, what happens to people with that gene? http://www.youtube.com/watch?v=XuUpnAz5y1g&feature=related Cloning A clone is an organism that is a perfect copy of another organism Clones can happen in nature Identical twins Some organisms (like bacteria) “clone” themselves through asexual reproduction Cloning Cloning mammals is really hard we used to think it was impossible Dolly the sheep – 1996 Making a Clone: 1. A cell is taken from the donor. 2. An egg is taken from another adult animal. 3. The egg has all of its genetic material (DNA) removed. 4. The genetic material (DNA) is taken from the donor cell and placed into the now-empty egg. 5. The egg with the donor’s genetic material is placed in a foster mother where it will divide and grow into a baby. This is the clone, a perfect copy of the original donor. Problems with Cloning: Most attempts to clone have failed Of the attempts that succeed, most have health problems Large offspring syndrome (LOS) – clone babies tend to be larger than other babies of their species the enlarged organs cause health problems Gene expression – a clone may not express the same genes as the original, even if they are genetically identical Problems with Cloning Clones tend to have weak immune systems they are more vulnerable to disease Cells tend to have more trouble dividing properly the older an organism is if you take an “old” cell from an older organism and grow a younger organism (clone) from it, there will be even more dividing problems Therefore, it is currently illegal to clone humans in the United States What is Gene Therapy? The transplantation of normal genes into cells in place of missing or defective ones in order to correct genetic disorders. Gene Therapy Still in the theory/testing stages 1. A person is tested for faulty genes 2. A person with a healthy copy of the gene donates some cells and has the healthy copy cut out using restriction enzymes 3. The healthy copy is loaded into a vector (carrier), usually a virus. 4. The sick person is infected with the virus, which will inject the gene into their cells, hopefully “patching up” the problem. http://www.youtube.com/watch?v=CkWep1Z0gCw Diseases that can be cured with Gene Therapy 1. Cystic fibrosis = causes respiratory tract to produce a thick mucus (instead of the normal thin, watery kind) that can clog passageways and lead to suffocation. People with CF usually die by 30 2. Severe Combined Immunodeficiency (SCID) = affects two kinds of infection fighting cells, leaving patients susceptible to diseases. Women are usually only carriers, men are usually the ones affected. Problems with Gene Therapy Very expensive to research and test We don’t know what the side effects will be We still don’t understand everything about genes how do we know this will fix the problem? Stem Cell Research Stem cells are a “blank slate” they can become any kind of cell through differentiation ex. Heart cell, lung cell Two main kinds: Adult stem cells = come from adults Embryonic stem cells = come from embryos grown in the lab Stem Cell Research Scientists prefer to work with embryonic stem cells Easier to identify, isolate, and harvest More of them than adult stem cells Grow more quickly and easily Easier to manipulate Problems with Stem Cell Research Financial It is very expensive Ethical Destroying embryos Stem cells can cause cancer if they divide out of control Why use embryonic when adult cells are available? Adult stem cells avoid problem of immune rejection