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Recombinant Dna And Polymerase Chain Reaction PowerPoint Presentation

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Slide 1 - RECOMBINANT DNA AND POLYMERASE CHAIN REACTIONS
Slide 2 - DNA – Double Helix Structure Each spiral strand is composed of a sugar phosphate backbone and attached bases 4 Bases: Adenine (A), Guanine(G), Cytosine (C), and Thymine (T). Form Base Pairs; A with T and C with G in the complementary strand via hydrogen bonding (non- covalent) The strands can be cut by restriction enzymes, e.g. ECOR1 What do we already know
Slide 3 - Bacteria are often used in biotechnology as they have plasmids A plasmid a circular piece of DNA that exists apart from the chromosome and replicates independently of it. Bacterial Structure
Slide 4 - Fill in the blanks of the worksheet
Slide 5 - DNA that has been cut from one strand of DNA and then inserted into the gap of another piece of DNA that has been broken. The host DNA is often a bacterial cell such as E coli. The purpose of splicing the gene into the host DNA is to produce many copies of it. As bacteria reproduce in a very short time it is possible to make millions of copies of the gene fairly quickly. What is Recombinant DNA?
Slide 6 - The required gene e.g. Insulin, is cut from the DNA using a restriction enzyme. A circular piece of DNA, called a plasmid, is removed from the bacterial cell and is cut open using the same restriction enzyme. The cut out human gene is then mixed with the bacterial plasmids in a test tube. Because they have been cut with the same enzyme, the cut ends of the plasmid and the end of the human gene match. Often called ‘sticky ends’ The enzyme DNA ligase is used to stick the ends together. How do we make it?
Slide 7 - ppt slide no 7 content not found
Slide 8 - Re-Introducing the Plasmid Back - TRANSFORMATION Now the plasmids that contains the introduced gene (recombinant DNA) need to be reintroduced into the bacteria so they can multiply and make more of the gene. Can be done by combining them in a test tube with CaCl2. The high concentration of calcium ions makes the membranes of the bacteria more porous. This then allows the plasmids to move into the bacterial cells. Not all bacteria will take up a plasmid and this is why the monitoring must happen.
Slide 9 - How do we know which bacteria have the gene? It is necessary to isolate the host bacteria that contain the gene that has been spliced as only want the recombinant DNA By having a gene on the same plasmid that gives resistance to an antibiotic, the other bacteria can be removed by culturing the bacteria in a medium that contains the antibiotic. The bacteria containing the resistance to the antibiotic will survive and the others will be killed by the antibiotic.
Slide 10 - Selection of Altered Cells Antibiotic resistance gene used to identify recombinant cells
Slide 11 - http://www.sumanasinc.com/webcontent/animations/content/plasmidcloning.html ANIMATION – RECOMBINANT DNA
Slide 12 - Plasmids will not work as well in eukaryotic organisms like plants and animals Other methods need to be used to insert the DNA Viral vectors can be used for animal cells. The virus can ‘inject’ their DNA into an animal host cell. Can we use this technique on all cells?
Slide 13 - Gene Gun can be used to insert genes into plant cells http://www.hort.purdue.edu/hort/courses/HORT250/animations/Gene%20Gun%20Animation/Genegun1.html GENE GUN
Slide 14 - Biotechnologists are Problem Solvers! Diabetics having reactions to porcine/animal insulin Wheat crops being attacked by insects People sick with cystic fibrosis All these can be fixed by recombinant DNA!!! On a Flow Chart show the steps involved in making recombinant DNA for a desired gene. From cutting of the gene to the final product (this may involve the delivery method)
Slide 15 - Now we have made the gene – how do we get lots of copies??
Slide 16 - E.Coli Plasmid is cut with the same restriction enzyme used to cut the insulin gene Insulin gene is cut from a pancreatic cell DNA using a specific restriction enzyme insulin - Bacterial cells when supplied with required polypeptides or proteins, the colonies will produce insulin E.g Vaccines- The plamids are isolated from the e.coli cells, the genes are then amplifyed via PCR and used to create inactivated viruses for vaccines
Slide 17 - Useful Properties of DNA The complementary strands of DNA can be separated and re-associated by heating and cooling One strand of DNA specifies the sequence of the other strand
Slide 18 - Polymerase Chain Reaction Used to make more copies of DNA from a tiny DNA sample http://www.sumanasinc.com/webcontent/animations/content/pcr.html
Slide 19 - Polymerase Chain Reaction (PCR) Amplifies DNA Primers specify what DNA is copied
Slide 20 - PCR Amplifies DNA Diagnosis Epidemiology Genetic engineering
Slide 21 - Flow Chart of PCR
Slide 22 - Production of Insulin Making recombinant vaccinations Making food crops with immunity to insects Forensic Crime scene analysis – DNA profiling Real World Applications
Slide 23 - Other Issues Ethical issues related to “cloning” of human genes How will genetically engineered organisms affect environment? Spread of genes to other organisms? Who will decide?
Slide 24 - ppt slide no 24 content not found
Slide 25 - This powerpoint was kindly donated to www.worldofteaching.com http://www.worldofteaching.com Is home to well over a thousand powerpoints submitted by teachers. This a free site. Please visit and I hope it will help in your teaching
 

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