When many of us think of biology the smallest we get is the cellular level. In Dr. Fernández-Robledo’s lab we get even smaller than that, down to the genetic level. Much of Dr. Fernández-Robledo’s work is concerning the oyster parasite Perkinsus marinus. This parasite causes the “Dermo” disease in oysters which causes the cells in the oysters to essentially dissolve. Because of this, an understanding of Perkinsus is important economically as well as to foster a greater understanding of marine parasite ecology. This is just one model species though. We work with many species of microorganisms mapping and manipulating their genomes to better understand where different genes are and how they do what they do.
I will be the first to admit much of my first conversation with Dr. Fernández-Robledo flew over my head. It was not until after two or three times of talking about my project and making notes of all the words I didn’t know to look up later that I could explain my project to my classmates. At this point to my best understanding I am attempting to transfect two species of marine protists with a couple different plasmids using eletroporation.
A few weeks ago I (who have never taken a microbiology class) would have recognized perhaps half of the words in the previous sentence. Now I have my own little lab bench area and I’m up to about 75% understanding of the words used when talking about my research now. The learning curve here is pretty steep, but because we’re so immersed in the program it’s fairly easy to catch up. I’ll do my best to explain my project now and apologize to Dr. Fernández-Robledo for any inaccuracies.
Transfection is the process of putting DNA (Deoxyribonucleic acid, the instruction booklet for life) into eukaryotic cells (these are the cells of organisms that have a nucleus and membrane bound organelles, organisms like humans, trees, and even single celled algae).
Protists are eukaryotes that aren’t animals, plants, or fungi. The ones I am using are Emiliania huxleyi and Chrypthecodinium conhii, a photosynthesizing coccolithophore (they make calcium carbonate shells) and a dinoflagellate respectively.
Plasmids are circles of DNA that are produced by bacteria and can be used in other organisms to identify where in the organism is expressing a gene by binding to a section of the receiving organism’s genome. For my project I am using plasmids that have an indicator attached to them, so when they are incorporated into the receiving organism they glow with either red or green fluorescence (depending on the plasmid) so we can tell if it worked. So far I’ve extracted plasmids from the bacteria we were using to reproduce the genes. This has been my first foray into lab work so I still have a lot to learn about the processes, but under the watchful gaze of Dr. Fernández-Robledo I managed to bumble my way through that first step in my project.
Electroporation is the way in which we will be doing the transfection. Electroporation works by essentially putting the cells and plasmids you want combined in a conductive solution and then zapping it. Then, ideally, the cells’ membrane will open up and the plasmids will get in there and become incorporated. I will be trying to zap the cells with a few different patterns of impulses.
In addition to my work with Dr. Fernández-Robledo, I also have the opportunity to interact with scientists who do work ranging from kelp herbivory ecology to oceanic modeling. Meeting people with such varied backgrounds provides a great resource to learn about not only the work that is being done in the ocean, but ways in which one could enter the field.
Duncan Coles is a Colby College student in Bigelow Laboratory for Ocean Science’s semester-in-residence program. This intensive research experience is focused on ocean science within a changing global climate.