Collaborative Endeavors and Expertise in Various Fields
Dustin Ebert
Sufia Munim
Sudeep Shakya
Create an E. coli that glows green
while smelling like wintergreen in
presence of salicylic acid.
The aim of the project is to…
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
1.
To isolate the Biobricks parts necessary for the
project. Next step is to add and ligate parts
together in four different experimental
arrangements.
a.
Promoter–Wintergreen gene
b.
Promoter-GFP
c.
Promoter-Wintergreen-GFP
d.
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.
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:
Wintergreen
gene
Methyl salicylate
Induced
promoter
GFP
Regular
Promoter
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.
•
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.
Promoter
Wintergreen gene
◦
Cut promoter out and add to wintergreen plasmid
Promoter
GFP gene
◦
Cut promoter out and add to GFP plasmid
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.
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.
Embrace the collective wisdom and diverse backgrounds of Dustin, Ebert, Sufia, Munim, and Sudeep. Explore innovations, insights, and collaborations in different domains. Unleash the power of teamwork and shared knowledge to spark creativity and drive positive change. Together, they bring a plethora of experiences and skills to the table, paving the way for exciting partnerships and impactful projects that transcend boundaries and inspire others.
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Presentation Transcript
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.
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
To isolate the Biobricks parts necessary for the project. Next step is to add and ligate parts together in four different experimental arrangements. a. Promoter Wintergreen gene b. Promoter-GFP c. Promoter-Wintergreen-GFP d. Promoter-GFP-Wintergreen gene 1. The biobricks would all end up arranged on a pSB1A3 plasmid except for (b) which would be on a pSB1A2 plasmid.
2. Experiment with different concentration of activity. 3. If time permits, we will look for a promoter that allows this arrangement: gene L-Arabinose to examine the promoter Methyl salicylate Induced promoter Regular Promoter Wintergreen 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.
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.
PromoterWintergreen gene Cut promoter out and add to wintergreen plasmid Promoter GFP gene Cut promoter out and add to GFP plasmid
PromoterWintergreenGFP 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.
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.
