Module 2: Expression EngineeringModule 2: Expression Engineering20.109Lecture 2October 16th, 2007Cite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY].Public Understanding?http:// www.ardenmoore.com/pets/dna-diet.htmlImage removed due to copyright restrictions. From “The DNA Diet,” AKC Gazette, April 2006:“Scientists are seeking the recipes for dog foods that can attack illness at the genetic level”Cite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY].Public Understanding?Images removed due to copyright restrictions. Website screenshots from http://www.thednadiet.com, advertising a service using DNA testing to develop a personal diet plan.Cite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY].Public Understanding?Images removed due to copyright restrictions.Photo of anti-GMO protesters from http://www.gmfreeireland.orgCartoon of “mad scientist” characterEditorial cartoon about fast-track bill to allow use of genetically modified food ingredients without labelingCite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY].Public Understanding?Image removed due to copyright restrictions.Cartoon of “mad scientist”character.Cite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY]. Text removed due to copyright restrictions.Excerpt from "Lawmakers, Administration Clash over Science Unding." by Aliya Sternstein. National Journal's Technology Daily, February 14, 2007.http://www.govexec.com/story_page.cfm?articleid=36137&ref=rellink (accessed May 7, 2008)Expression Engineering ExperimentDay 1Day 2Day 3Day 4Day 5Day 6Image of glowing luciferasein a microcentrifuge tube, removed due to copyright restrictions.RTFigure by MIT OpenCourseWare.Image credits: Day 1 - Courtesy of Arkitek Studios. Used with permission. Day 2 - Courtesy of The Exploratorium. © The Exploratorium, http://www.exploratorium.edu. Day 4 – Figure by MIT OpenCourseWare. Day 5 – Dr. Natalie Kuldell. Day 6 – Courtesy of NIH.Cite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY].Expression Engineering ExperimentLecture 1• intro to cell culture• intro to gene exp’n/RNAiLecture 2• transfection• luciferaseLecture 3• off-target/nonspecific RNAiLecture 4• Writing lecture(Neal Lerner)Lecture 5• measuring gene express’nLecture 6• microarray analysis (Rebecca Fry)Lecture 7• high throughput technologies or RNAi applications (no lab)Lecture 8• review of your dataCite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY].DNA on the move: nature’s waysFigure by MIT OpenCourseWare.Phage infects Occasional phagepackages host DNAHybrid phageinfects new cellHost may incorporate foreign DNACite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY].DNA on the move: nature’s ways1. Viral transductionhttp://blogs.nature.com/news/blog/2006/10/ancient_human_virus_resurrecte.htmlCite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY]. Text and image removed due to copyright restrictions.Excerpt from: Pearson, Helen. "Ancient Human Virus Resurrected."Nature (31 October 2006). DOI:10.1038/news061030-4.DNA on the move: nature’s ways1. Viral transduction2. ConjugationFigure by MIT OpenCourseWare.Mobile plasmidPilusBacteriumChromosomeDonorRecipientRelaxosome TransferosomeNew donor New donorABCDCite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY].DNA on the move: nature’s ways1. Viral transduction2. Conjugation3. TransformationJonathan M. Solomon and Alan D. Grossman “Who's competent and when: regulation of natural genetic competence in bacteria” Trends in Genetics (1996) 12(4):150Courtesy Elsevier, Inc., http://www.sciencedirect.com. Used with permission.Cite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY].DNA on the move: in the lab1. Viral transduction2. Conjugation3. TransformationchemicalelectricalCaCaPPPPFigure by MIT OpenCourseWare.Courtesy of Zephyris.Cite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY].CaCaPPPPFigure by MIT OpenCourseWare.DNA on the move: in the lab1. Viral transduction2. Conjugation3. TransformationchemicalelectricalmicroinjectionbiolisticsCourtesy of Zephyris.Cite as: Natalie Kuldell. Course materials for 20.109 Laboratory Fundamentals in Biological Engineering, Fall 2007. MITOpenCourseWare (http://ocw.mit.edu), MassachusettsInstitute of Technology. Downloaded on [DD Month YYYY]. Image removed due to copyright restrictions.Image removed due to copyright restrictions.See http://firstivf.net/laboratory_tour.htm.Nature Biotechnology (2007) 25: 1149 - 1157 Mechanisms and