Lab 13: Genetic Engineering PDF

Lab 13: Genetic engineering Announcement Discovery Coast assignment and Presentations!! Final exam: Lab 11, 13, 1, 5 NO Biodiversity :) Final class is super mega extra mandatory do not miss it! We’re working with E.coli today so put the water away pls What is Genetic Engineering? Geneti...

Lab 13: Genetic engineering Announcement Discovery Coast assignment and Presentations!! Final exam: Lab 11, 13, 1, 5 NO Biodiversity :) Final class is super mega extra mandatory do not miss it! We’re working with E.coli today so put the water away pls What is Genetic Engineering? Genetic Engineering is a way to modify an organism's genetic material by introducing new DNA Recombinant DNA means combining DNA from different sources to create new genetic material Example: Producing insulin for diabetes patients. The human gene for insulin is inserted into bacteria, which then produce insulin we can collect and use.  Sickle Cell Inheritance A person can be a carrier (or have the trait) or have the disease NOT sex-linked Reminder: Understanding DNA and Genes DNA: Contains genetic instructions, using four bases (A, T, G, C). Genes: Segments of DNA that contain instructions for making proteins. Proteins: Molecules that perform various functions in the body, made by reading gene sequences. Protein Synthesis Overview Protein Synthesis: Turning genes into proteins. 1. Transcription: DNA is transcribed into mRNA. 2. Translation: mRNA is translated at the ribosome to make a protein. Key Concept: DNA sequence determines the order of amino acids in a protein, defining its function. Investigation 1 - Protein Synthesis Activity Use the puzzle to simulate protein synthesis. FOLLOW THE DIRECTIONS IN YOUR LAB MANUAL!! Fig 13-3 directions to read codon chart What is a Plasmid? Definition: A plasmid is a small, circular DNA molecule found in bacteria that replicates independently of chromosomal DNA. Use in Genetic Engineering: Plasmids are used as vectors to carry foreign genes, such as the insulin gene, into bacteria. Antibiotic Resistance: Plasmids often carry genes that provide resistance to antibiotics, used as markers to identify successful transformations. Restriction Enzymes Restriction enzymes are proteins that cut DNA at specific sequences. Sticky Ends: These cuts create overhanging sequences called "sticky ends" that help insert new DNA into plasmids. EcoR1: A commonly used enzyme that makes cuts ideal for inserting foreign DNA without disrupting important genes. Investigation 2 - Genetic Engineering Simulation Activity: Cut open a plasmid and insert the human insulin gene using restriction enzymes. Goal: Open up a plasmid and insert a human gene (insulin) Set up: Remember last time? Cut and tape in correct order. Green sheet for human DNA. Yellow sheet is the bacterial plasmid DNA Steps: Two restriction enzymes, make notes of where Would cut before cutting. Choose the correct restriction enzyme then make Your cuts! ***Only 1 enzyme cuts both DNA strands*** *** Gray areas MUST BE INTACT*** (this is where the resistance to the antibiotic is) E. coli Overview Definition: E. coli (Escherichia coli) is a common bacterium found in the intestines of humans and animals. Role in Lab: E. coli is often used in genetic engineering because it is easy to grow and can take up foreign DNA. Why E. coli?: It replicates quickly, making it useful for producing large quantities of a protein of interest after transformation. Heat Shock Process Purpose: To help plasmids enter bacterial cells. Process: Bacteria are briefly exposed to heat, which makes their cell membranes more permeable so the plasmid DNA can enter. Importance: This is a key step in genetic transformation to introduce new genes into bacteria. Investigation 3 - Transformation of E. coli Activity: Introduce the luminescent plasmid into E. coli using heat shock, then plate the bacteria. Background: Luciferin and Luciferase: Genes responsible for the glow, originally from fireflies. Ampicillin Resistance: Allows us to identify bacteria that successfully took up the plasmid. Expected Results: Only transformed bacteria will grow on ampicillin plates, and they will glow.