AP Bio: Wednesday, 2.16.11 Planning Lab 6A; Mutations

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AP Bio: Wednesday, 2.16.11 Planning Lab 6A; Mutations. Homework: Study for Test on Molecular Genetics Do Now: Get a copy of Lab 6A: Transformation Todays Goals: Design a procedure for genetically engineering E. coli cells to glow. Describe the effects of different types of mutations. - PowerPoint PPT Presentation

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<ul><li><p>Homework:Study for Test on Molecular GeneticsDo Now:Get a copy of Lab 6A: TransformationTodays Goals:Design a procedure for genetically engineering E. coli cells to glow.Describe the effects of different types of mutations.Agenda:Mini-Lecture: Intro to Lab 6A TransformationExplanation of Lab ReportIn Lab Groups:(1) Use the colored cards to make a flow chart and then a procedural list for completing this lab.(2) Complete the Computer-Based Mutations Lab. Print and hand to me when finished.</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>AP Biology Lab 6:Genetic EngineeringviaBacterial TransformationMaking E. coli glow like jellyfishAmy Dickson, Prospect Hill Academy Charter SchoolAll images by Christine Rodriguez and Amy Dickson</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>GOALS OF THIS LAB PROJECT: Make E. coli bacteria glow like jellyfishBy inserting the GFP (green fluorescent protein) gene from a jellyfish into a bacterial plasmid Control when the bacteria express this proteinBy connecting the GFP gene to an on/off switch that causes it to be expressed only in certain environments</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>WHY SHOULD WE DO THIS?Genetic Engineering is now widely used: Bacteria that produce human insulin Corn that produces insecticide Rice that produces extra vitamin A Goats that produce spider silk</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>WHY SHOULD WE DO THIS?To SEE the Central Dogma in action:</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>WHY SHOULD WE DO THIS?To SEE the Central Dogma in action:To understand how gene expression is regulated - how cells (and the scientists who manipulate them) control when genes are turned on/off.onXTrait</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>A BIT OF BACKGROUND ON GENE REGULATIONPromoter: A short DNA sequence upstream of a gene where RNA pol. binds to start transcription Serves as the on/off switch for the gene blocking it turns the gene offWhy do this?Making proteins only when needed saves energy and materials</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>A BIT OF BACKGROUND ON GENE REGULATIONpromotergenes for arabinose-digesting enzymesgenes not expressedAra repressor (active)Arabinose sugar binding siteArabinose sugar (inducer)genes expressed!Example: Arabinose is a sugar that bacteria can digestBut no need to make enzymes unless arabinose is aroundNormal condition: Promoter blocked by Ara repressorIn presence of arabinose: repressor is inactivated; gene is turned on</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>QUICK REVIEWPromoter - </p><p>Plasmid - </p><p>Transformation - </p><p>a process in which bacteria take up DNA from their environment a small, circular piece of bacterial DNA that is not part of the chromosome an on/off switch for a gene- can be triggered by electric shock or heat shock </p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>STARTING MATERIALSBacterial chromosomeE. coli cells sensitive to antibiotics cant glow competent - able to be transformed</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>STARTING MATERIALSPlasmid containing:</p><p> Ampicillin resistance gene (always expressed)</p><p> Ara promoter - turned on in the presence of arabinose </p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>STARTING MATERIALSJellyfish DNAGFP = Green Fluorescent Proteinglows under UV light</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>Jellyfish DNASTARTING MATERIALSE. coli cellsPlasmid</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>END RESULTRecombinant Bacteria</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>HOWEVERthings are actually a bit more complex.AmpRArapromoterGFPpGLO plasmid</p><p>Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College.</p><p>YOUR TASK:Design an experimental procedure for genetically engineering glowing bacteria.Goals to consider:#1 - Make recombinant bacteria#2 - Select for only the recombinant bacteria#4 - Establish a control for your experiment to demonstrate that its the plasmid that causes ampicillin resistance and the ability to glow.#3 - Make the recombinant bacteria glow only when we want them to.</p></li></ul>