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BioBuilder : Engineering Cells

BioBuilder : Engineering Cells . James Dixon Sharon High School Rebekah Ravgiala Tyngsborough High School Aaron Mathieu Acton Boxboro High School. Natalie Kuldell. Our Goals. Can we grow houses? Not yet, but synthetic biologists have…. Engineered bacteria to change color

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BioBuilder : Engineering Cells

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  1. BioBuilder: Engineering Cells James Dixon Sharon High School RebekahRavgiala Tyngsborough High School Aaron Mathieu Acton Boxboro High School

  2. Natalie Kuldell

  3. Our Goals

  4. Can we grow houses? Not yet, but synthetic biologists have…

  5. Engineered bacteria to change color based on light Evolution didn’t do this! Engineers did… And they have…

  6. Abstraction Hierarchy

  7. Parts

  8. A device A system

  9. link

  10. teachers

  11. If given ONPG* as a substrate (*o-nitrophenyl-β- D-galactoside) LacZ will produce Galactose o-nitrophenol, Which is yellow! An indicator of enzyme activity • Promoter: • Strong • Medium • Weak • RBS: • Strong • Medium • Weak LacZ ORF

  12. Computer aided design (CAD) generated model:

  13. We can use these slide bars to alter the system

  14. And generate data like this…

  15. Modeling the cell with electronics…

  16. A Bread Board Model:

  17. “BioBuilding” Professional Development Workshop @MIT

  18. Synthetic Biology and Engineering Education Synthetic Biology and Engineering Education

  19. The BioBuilder curriculum

  20. Synthetic Biology“…aims to apply standardized engineering techniques to biology and thereby create organisms or biological systems with novel or specialized functions to address countless needs.” (Science 33:6047, 9/2/11) Eau that Smell Competing Designs for BioBuilding! MIT iGEM 2006 Rebekah Ravgiala, Tyngsborough High School MABT, 3/10/12

  21. ABS Plastic Device “DNA” Input Output Part Abstraction Hierarchy for Biological Systems Analogy Black box functions System Rebekah Ravgiala, Tyngsborough High School MABT, 3/10/12

  22. Design Test The focus of this lab Build Engineering & Design CycleWhat’s going on in this lab? Rebekah Ravgiala, Tyngsborough High School MABT, 3/10/12

  23. Potential Learning Objectives • What is synthetic biology? • Synthetic biology vs. genetic engineering. • Investigate population growth curve of bacteria. • Practice standard microbiology methods. • Measure the growth of a bacterial population. • Define and properly use synthetic biology terms (parts, device, system) • Define and properly use molecular genetics terms (promoter, RBS, ORF, Terminator, Plasmid) *Highlighted are the LO focused on… Rebekah Ravgiala, Tyngsborough High School MABT, 3/10/12

  24. Background MIT students (iGEM 2006) designed Eau d’coli, E. coli that smell like bananas when their population is in the stationary phase. System Components 1. Stationary phase promoter 2. BGD (a banana smell device) a. RBS b. ORF (ATF1 enzyme & terminator sequence) *The ATF1 enzyme converts isoamyl alcohol to isoamyl acetate, the molecule that gives bananas their characteristic smell. Rebekah Ravgiala, Tyngsborough High School MABT, 3/10/12

  25. Challenge: Evaluating Competing Designs Strain 1-1.(+ control) Original MIT Device Strain 1-2. Original MIT Device + Inverter Strain 1-3. Log Phase Promoter + BGD Strain 1-4. (- control) No Smell Generating Device Challenge Objectives: 1. Grow these bacterial populations 2. Test for the banana smell of the population from lag to stationary phase. 3. Measure the density of the culture by using a Spec 20 OR the McFarland Turbidity Standards 4. Compare the banana smell to dilutions of banana extract standard. Rebekah Ravgiala, Tyngsborough High School MABT, 3/10/12

  26. Workflow Protocol A Prepared liquid cultures of each strain. a. Bacterial strains have been transformed with plasmids corresponding to 1-1, 1-2, 1-3, 1-4 b. They may arrive as slants or on Petri dishes or as liquid cultures c. Prepare stock growth media (LB, amp, isoamyl alcohol) d. Use 75ml of stock and add liquid cultures Strain 1-1 Strain 1-2 Strain 1-3 Strain 1-4 Student Lab Protocol and Instructional Videos Rebekah Ravgiala, Tyngsborough High School MABT, 3/10/12

  27. T0 T0 T0 T0 T0 T0 T0 T0 Strain 1-1 Strain 1-2 Strain 1-2 Strain 1-3 Strain 1-4 Workflow Protocol A 1. Prepared liquid cultures of each strain. For each strain, place a 25ml sample in the refrigerator before it has time to incubate (lag phase). Rebekah Ravgiala, Tyngsborough High School MABT, 3/10/12

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