Chapter 20: Terms to Know 1. Genetic engineering 2. Biotechnology 3. Recombinant DNA 4. Gene cloning 5. Restriction enzymes 6. Sticky ends 7. DNA ligase

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Text of Chapter 20: Terms to Know 1. Genetic engineering 2. Biotechnology 3. Recombinant DNA 4. Gene cloning...

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Chapter 20: Terms to Know 1. Genetic engineering 2. Biotechnology 3. Recombinant DNA 4. Gene cloning 5. Restriction enzymes 6. Sticky ends 7. DNA ligase 8. Cloning vector 9. Nucleic acid hybridization 10. Genomic library 11. cDNA library 12. PCR 13. Gel electrophoresis 14. Southern blotting 15. DNA microarray assays 16. SNPs 17. RFLPs 18. Stem cells 19. Gene therapy 20. GMO (genetically modified organism) Slide 2 What You Must Know: The terminology of biotechnology. The steps in gene cloning with special attention to the biotechnology tools that make cloning possible. The key ideas that make PCR possible. How gel electrophoresis can be used to separate DNA fragments or protein molecules. Slide 3 Genetic Engineering : process of manipulating genes and genomes Biotechnology : process of manipulating organisms or their components for the purpose of making useful products. Slide 4 Recombinant DNA : DNA that has been artificially made, using DNA from different sources eg. Human gene inserted into E.coli Gene cloning : process by which scientists can product multiple copies of specific segments of DNA that they can then work with in the lab Slide 5 Tools of Genetic Engineering Restriction enzymes (restriction endonucleases): used to cut strands of DNA at specific locations (restriction sites) Restriction Fragments: have at least 1 sticky end (single-stranded end) DNA ligase : joins DNA fragments Cloning vector : carries the DNA sequence to be cloned (eg. bacterial plasmid) Slide 6 Using a restriction enzyme (RE) and DNA ligase to make recombinant DNA Slide 7 Gene Cloning Slide 8 Applications of Gene Cloning Slide 9 Techniques of Genetic Engineering Slide 10 Transformation : bacteria takes up plasmid (w/gene of interest) Nucleic acid hybridization : used to track gene of interest PCR (Polymerase Chain Reaction): amplify (copy) piece of DNA without use of cells Gel electrophoresis : used to separate DNA molecules on basis of size and charge using an electrical current (DNA + pole) Southern blotting : used to find a specific human gene DNA microarray assays : study many genes at same time Slide 11 PCR (Polymerase Chain Reaction) : amplify (copy) piece of DNA without use of cells Slide 12 Nucleic Acid Hybridization: find and track gene of interest Slide 13 Gel Electrophoresis: used to separate DNA molecules on basis of size and charge using an electrical current (DNA + pole) Slide 14 Slide 15 Southern Blotting: used to find a specific gene Slide 16 Microarray Assay: used to study gene expression of many different genes Slide 17 DNA microarray that reveals expression levels of 2,400 human genes Slide 18 Cloning Organisms Nuclear transplantation : nucleus of egg is removed and replaced with nucleus of body cell Slide 19 Nuclear Transplantation Slide 20 Problems with Reproductive Cloning Cloned embryos exhibited various defects DNA of fully differentiated cell have epigenetic changes Slide 21 Stem Cells Stem cells : can reproduce itself indefinitely and produce other specialized cells Zygote = totipotent ( any type of cell) Embryonic stem cells = pluripotent ( many cell types) Adult stem cells = multipotent (a few cell types) or induced pluripotent, iPS (forced to be pluripotent) Slide 22 Embryonic vs. Adult stem cells Slide 23 Using stem cells for disease treatment Slide 24 Applications of DNA Technology 1. Diagnosis of disease identify alleles, viral DNA 2. Gene therapy alter afflicted genes 3. Production of pharmaceuticals 4. Forensic applications DNA profiling 5. Environmental cleanup use microorganisms 6. Agricultural applications - GMOs Slide 25 Gene therapy using a retroviral vector Slide 26 Pharm animal: produce human protein secreted in milk for medical use Slide 27 DNA Fingerprinting Slide 28 RFLPs (rif-lips) Restriction Fragment Length Polymorphism Cut DNA with different restriction enzymes Each person has different #s of DNA fragments created Analyze DNA samples on a gel for disease diagnosis Outdated method of DNA profiling (required a quarter-sized sample of blood) Slide 29 RFLPs Disease Diagnosis Slide 30 STR Analysis STR = Short Tandem Repeats Non-coding DNA has regions with sequences (2- 5 base length) that are repeated Each person has different # of repeats at different locations (loci) Current method of DNA fingerprinting used only need 20 cells for analysis Slide 31 STR Analysis Slide 32 Slide 33 Slide 34 Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique What are the main tools or materials involved? Applications: What is this being used for?

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