Dr. Gong’s lab studies fluorescent proteins and their application in neural imaging. Fluorescent proteins are a type of protein originally discovered in jellyfish that have now been modified to display an array of colors and can be used in many different studies. In the Gong lab the goal is to optimize the brightness and folding characteristics of fluorescent proteins so the brain can be imaged at a higher degree of detail so more of the function can be understood. To accomplish this a virus will insert the DNA for two fluorescent proteins linked to a calmodulin domain and a M13 muscle domain. This combination is extremely sensitive to calcium concentration, which fluctuates greatly in the brain, so depending on the amount of calcium near the proteins the color they produce in an image will change. This enables neural networks to be imaged at one moment so the interactions between a large group of neurons can be observed at once, leading to a much greater understanding of the brain’s function.
My work in the lab focuses on the optimization of the mNeonGreen fluorescent protein. I am primarily focusing on two things to accomplish during my summer, one wet lab based, and the other programming based. For the wet lab portion I am creating different versions of the mNeonGreen protein linked to a consistent mRuby protein that will be used as a control. This process includes primer design, PCR, and a few other processes that will create bacteria colonies that will produce the proteins. The different versions I am using are a circularly permuted version of the protein and other versions that have slight mutations that will hopefully improve the brightness and expression of the protein.
Once the different colonies are created I will analyze them using a program that I am writing on Matlab. After each colony is grown for 18 hours I take a picture of the plate so the fluorescence of the proteins can be analyzed. The goal of the program is to determine the location of all of the colonies and then find the colony that has the highest ratio of green to red fluorescence. The high ratio of green will indicate that the version of the protein was particularly bright and effective compared to the controlled red.
I’m excited to see the results that will come from the summer!
