Agne Jakubauskaite ’13 has come full circle in the course of her undergraduate research of the newly discovered gene TBhR. Jakubauskaite, a biology concentrator, spent the summer of 2011 learning the ins and outs of protein expression and synthesizing and has now passed on those skills to Jessica Li ’14, a biology concentrator and Olusegun Ogunwomoju ’15.
After teaching Ogunwomoju and Li the fundamentals of this year’s lab work, Jakubauskaite left for Harvard University where she has been accepted into the prestigious Stem Cell Institute Summer Undergraduate Research Program. Jakubauskaite started her work at Harvard in the beginning of June, and she has been assigned to the Kim Lab in Boston Children's Hospital. Ogunwomoju and Li are now working independently from Jakubauskaite; however, her instruction has left the two well-prepared.
The TBhR gene is found in fruit flies and was originally discovered by Professor of Biology Herman Lehman, who oversaw Jakubauskaite’s research last year and is providing guidance this year as well. The newly discovered TBhR gene is similar to the MOX gene found in humans, and as such, its implications to the medical field are very significant. TBhR’s function is still yet to be determined, but given that it is so similar to MOX and is expressed in the development of the nervous system (specifically in the synthesis of neurotransmitters), the discovery of its purpose could lead to a greater understanding of the working of the central nervous system and its various disorders. According to Jakubauskaite, “abnormalities in the synthesis of neurotransmitters have been linked to many neurological diseases ranging from depression to Parkinson’s. Therefore, detailed understanding… might lead to a development of new treatments.”
In order to better determine TBhR’s function, Ogunwomoju and Li must synthesize the protein that it encodes and a complex process must then be undertaken to locate TBhR within the cell. Jakubauskaite has already made significant strides in completing this process, but now that she has left for her research at Harvard, the task falls to Ogunwomoju and Li.
This work, which is instrumental to Jakubauskaite’s senior thesis, involves tagging the colorless TBhR with a green protein. In order to visually expose TBhR, Ogunwomoju and Li first begin with a commercial plasmid containing TBhR and a green fluorescent protein plasmid (GFP). The TBhR and GFP plasmids are amplified by being inserted into bacteria in a process called transformation. When bacteria containing each of the two plasmids replicate, the plasmids are replicated as well, giving the team a greater pool of DNA to work with. After a large number of the bacteria are grown, they are then killed off and their DNA is extracted.
The extracted TBhR gene is then copied in a process known as polymerase chain reaction. Special enzymes are then used which recognize and cut the TBhR gene out of the commercial plasmid. Next, DNA ligase (essentially superglue for DNA) is used to paste the TBhR gene into the GFP plasmid. These new hybrid plasmids are then re-amplified via the same bacterial process and extracted once more. They can finally be inserted into eukaryotic cells which will express the TBhR-GFP hybrid protein in a glowing green light, allowing it to be located within the cell under a microscope.
This entire process is slow and is often elongated by even the smallest of errors. Checkpoints exist at many levels, and Jakubauskaite remarked that these checkpoints often take up more time than the synthesizing and extracting procedure itself. According to the students and Professor Lehman, the long process is made more bearable by the many small rewards which spring up along the way. Much of the chemistry itself occurs on an invisible scale, so when images of these proteins are finally visible in the cells, the feeling is gratifying.
Perhaps the most rewarding aspect of the project comes in the form of independence. Jakubauskaite remarked that summer research introduces a level of self direction not normally found in lab classes, meaning that every decision made by the students bears more weight and responsibility. According to Li, working in the lab as an undergraduate researcher allows students “to develop lab techniques and put to use classroom knowledge in a way that guided labs do not.”
Jakubauskaite is well on her way to being prepared for graduate studies in cell and molecular biology, and Li too hopes to someday pursue a career in biology research. Jakubauskaite enjoys recreational track and field and Ogunwomoju is also an avid recreational athlete. Li is a Hamilton Association for Volunteering Outreach and Charity site coordinator and also volunteers for Project SHINE.
Agne Jakubauskaite is a graduate of Panevezio Sauletekio School (Lithuania), Jessica Li is a graduate of Brooklyn Technical High School (N.Y.) and Olusegun Ogunwomoju is a graduate of the Hotchkiss School (Conn.)