The Engineering Baking Yeast to Fluoresce experiment determines yeast’s ability to accept the Green Fluorescent Protein (a gene from jellyfish that enables them them to fluoresce) and fluoresce in response to black or blue light. Synthetic biology and genetic design were fundamental assets in understanding and performing the insertion of foreign DNA into yeast and outlining the experimental procedure knowing the biological components involved in both organisms. In order to successfully complete this project, it was imperative to have a volume adjustable pipette, nitrile gloves, pipette tips, petri plates, inoculation loops, sterile water, YPD agar with and without G418, yeast, UV filter film, centrifuge tubes, and a glass bottle. Through the course of the lab, sterility was most pertinent in preventing contamination and spearheading the accuracy of the experimental results — especially since the lab was performed in a non-sterile environment (in a house). The yeast did glow as a result of the Green Fluorescent Protein (GFP).
In this lab, the goal was to make yeast fluoresce as a result of inserting GFP. The major elements in this experiment included 1) maintaining sterility 2) ensuring accuracy 3) following protocol and 4) being knowledgable about how to perform the experimental procedure. In maintaining sterility, replacing nitrile gloves every few hours, or after handling different fluids, can prevent contamination of ingredients. This is hand-in-hand with ensuring accuracy since accuracy is maintained through sterility and determines the learnings of the experiment. Without ensured accuracy, the lab is deemed insignificant in its findings. Following protocol drives the experiment by standardizing what’s necessary to achieve the end result. Knowledge about how to perform the experiment streamlines the entire procedure of the experiment.
The purpose of this experiment was to determine whether the GFP gene in jellyfish can produce a similar fluorescing effect when inserted in yeast.
If a GFP gene is inserted into yeast, then the yeast should glow, or fluoresce, in response to black or blue light because the effects of the gene in jellyfish are being implemented in the genome of yeast by leveraging synthetic biology.
- Volume adjustable pipette
- Pipette tips
- Nitrile gloves
- Petri plates
- Inoculation loops
- Sterile water
- YPD agar
- YPD agar with G418
- UV filter film
- Centrifuge tubes
- Glass bottle
- Make agar plates: Microwave agar and water in a glass bottle in 30 second increments until the solution is clear. Pour the solution onto petri plates. Leave the petri plates out for an hour with their lids on to allow the solution to solidify. Place the agar plates upside down in the fridge. Repeat the same procedure for agar with G418.
- Grow yeast on the agar plates: Dissolve a few grains of yeast in sterile water and shake the yeast and water together until they become a homogenous substance. Streak the yeast solution on both types of agar plates using an inoculation loop. Let the yeast grow by leaving it out in the open.
- Prepare the cell mixture: Add water to the expression plasmid and shake the water and freeze dried DNA until they are homogenous. Incubate the cell mixture in warm water. Add YPD media to the cell mixture and allow the mixture to incubate for 4–6 hours.
- Put everything together: Add the cell mixture to the YPD G418 agar plate. Use an inoculation loop to spread the mixture around. Flip the plate and store it in the fridge. Bring the orange film to your eyes and shine the blue light on the plates. If done correctly, the yeast should glow green.
Thoughts on bioluminescence:
On another note, bioluminescence, a concept similar to the one exemplified in the experiment, can be leveraged to mitigate the use of energy produced by modern lights and pose less harm to the environment. If enacting the same chemical reactions that occur in bioluminescent organisms in a light bulb can produce light, street lights and so many others can productively produce light without contributing to light pollution. Currently, light pollution contributes to nearly 2 billion tons of carbon a year.