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E.coli Tic Tacs

E.coli Tic Tacs. Dustin Ebert Sufia Munim Sudeep Shakya. The aim of the project is to… . Create an E. coli that glows green while smelling like wintergreen in presence of salicylic acid. Biobrick parts used… . Wintergreen gene ( BBa_J45004) Salicylic acid  methyl salicylate

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E.coli Tic Tacs

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  1. E.coli Tic Tacs Dustin Ebert SufiaMunim SudeepShakya

  2. The aim of the project is to… • Create an E. coli that glows green while smelling like wintergreen in presence of salicylic acid.

  3. Biobrick parts used… • Wintergreen gene (BBa_J45004) • Salicylic acid  methyl salicylate • Methyl salicylate smells like wintergreen • Green Fluorescent Protein (BBa_E0040) • Turns the recombinant E.coli green • Promoter (BBa_I0500) • induced by L-arabinose

  4. Project overview • To isolate the Biobricks parts necessary for the project. Next step is to add and ligate parts together in four different experimental arrangements. • Promoter–Wintergreen gene • Promoter-GFP • Promoter-Wintergreen-GFP • Promoter-GFP-Wintergreen gene The biobricks would all end up arranged on a pSB1A3 plasmid except for (b) which would be on a pSB1A2 plasmid.

  5. Project overview 2. Experiment with different concentration of L-Arabinose to examine the promoter activity. 3. If time permits, we will look for a promoter that allows this arrangement: Methyl salicylate Induced promoter Regular Promoter Wintergreen gene GFP This would prove that the wintergreen smell has been expressed in the culture. We might be able to find a methyl salicylate induced promoter in nature.

  6. Protocols • Since all the parts we are using are Biobricks , we won’t need to do PCR. • Use standard Biobricks restriction sites to cut out and arrange our parts.

  7. Protocols • PromoterWintergreen gene • Cut promoter out and add to wintergreen plasmid • PromoterGFP gene • Cut promoter out and add to GFP plasmid

  8. Protocols • PromoterWintergreenGFP • Cut promoter out and add to wintergreen plasmid • Cut GFP out and add it to our plasmid after the wintergreen plasmid. • PromoterGFPWintergreen • Cut promoter out and add it to GFP • Cut promoter-GFP product out and add to wintergreen plasmid.

  9. Testing the gene expression • Smell it. • If the promoter-wintergreen-GFP arrangement glows/green, we could assume that the wintergreen gene was expressed. • Try to find Methyl salicylate induced promoter.

  10. The end result

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