Hippo is one of the most important proteins in many pathways. Should there be a lack of it, mutations can occur, such as overgrown tissues, deformities and damaged skin (which resembles a large body of a hippopotamus, hence its name). Hippo can be a leading cause of cancers such as melanoma, breast cancer, brain cancer and testicular cancer. In a cell, hippo sends chemical messages towards the protein Yorkie. Yorkie is in the cells DNA, which is found in the nucleus. The purpose of Yorkie is to create more and more cells: the more proteins that are made, the bigger the organ will be. When hippo begins to notice a change it will send chemical messages to the Yorkie telling it to stop creating more cells. The overall organ size will get to the point where it’s too large for the fly’s body. If hippo is mutated, it is unable to send messages to the Yorkie and thus doesn’t tell it to stop producing cells.
Upon arrival, all the students partaking in the GTAC program reported to the Admin Manager to make their first official sign-in to the program (a process required at the beginning and end of each day). At 9:30am, everyone had gathered in the Valente room on the second floor of the building where Tony began the introductory stage of our 5-day program. He began with a basic outline of the week before assigning each of us a coloured card (red, yellow, green, blue), which determined our groups for the week.
After taking a seat at our assigned tables we began the ‘get-to-know us’ stage. The scientists involved in the program (Tony, KJ, Tom, Frazer and Claudia) introduced themselves, explained their field of expertise, how they got into said field and what drives their passion. Each group was then assigned a scientist and in our individual groups we had to figure out 6 things we all had in common with each other. We were then given a safety briefing before heading out to morning tea. Following on from our break, our groups were introduced to the blog aspect of the program in which we discussed the techniques, media and information that makes a good (and bad) blog. To conclude the day, our groups entered the Nossal laboratory, where we began the extraction of fly larvae DNA.
As Day 2 of the GTAC program commenced, we got straight into the practical components (now that we had completely finished our introduction to the weeks plans, safety components and team members). To kick the day off, each group got together and began their PCR set ups. We then heard from the scientist Cat who introduce herself (much like the others did yesterday) and gave us a rundown of her work and research conducted at the Peter MacCallum Institute. Following this, our group worked with DNA and PCR models, created a gel and used gel electrophoresis in the dark room and begin DNA sequencing in the laboratory.
Today was our 3rd day of the GTAC program in which we got to begin making our way through a 4-stage workshop. Each part of the workshop addressed different aspects of the overall project: fly genetics, SEM microscopy, bioinformatics, and blog preparation. Today our group got to do 3 of the 4: all except the SEM microscopy, which will be completed tomorrow.
Day 4 of the GTAC program commenced, our groups each completed their final stage of the rotations before working on their blogs for the afternoon. To conclude the day, all students partaking in the GTAC program attended the Peter Mac for a tour of one of their laboratories.
Today marked our final day of the GTAC program, we kicked off the morning with our final rotation, which consisted of modelling DNA sequences. Although our sequences were not returned in time for us to examine, we did get to analyse multiple genetic sequences on the computers in order to compare them with flies mutated with the hippo gene. Following on from our analysis, our groups spent the remainder of the day working on their blogs before submitting them and preparing our final presentation: which were to be presented to the other groups.
When we isolated the DNA from the fly larvae, we used Pre-Lysis Buffer to create the right conditions for the work and Proteinase K to break down proteins and the cell 'skeleton' so the nucleus was accessible. After breaking the nuclear membranes, a series of steps were undertaken for a purified DNA strand to be extracted.
Polymerase chain reaction (PCR)
During our GTAC experience, part of our research involved isolating a small portion of the DNA from the fruit flies, which meant the hippo gene had to be copied via PCR in order to obtain enough DNA for the sequencing. The PCR technique is used in molecular biology to make millions of copies of the DNA region from an initially small sample.
When analysing the hippo gene our group targeted 5 segments of the gene with some primers (short segments of DNA that complement the unique sequence on either side of the segment). In order to separate DNA strands from one another, the samples are heated in a high temperature before cooling down to a moderate temperature: this allows for the primers to bind to their target sequence. Finally, at moderate-high temperatures the DNA rebuilds itself and doubles its original amount.
When our group copied the barcoding region with the PCR, we used multiple ingredients (each with their own purpose): the primers physically bind to the DNA, Nucleotides (G, T, A, C) are used to duplicate the strands of DNA, Taq DNA polymerase adds the nucleotides together and speeds ups the process, the buffer maintains a consistent level and isolating the genomic DNA is used to duplicate certain sections of the DNA
This technique is used to separate DNA fragments according to their length (measured in base pairs, bp). DNA is a negatively charged molecule and when placed in an electric field with an applied current, it migrates to the positive electrode. The gel acts like a sieve, which means that smaller fragments can travel through it easier than the larger fragments.
SEM (Scanning Electron Microscope)
A scanning electron microscope (SEM) is a high-tech scientific piece of equipment that uses a beam of electrons in a vacuum (a space absent of air) in order to get high-quality images of a given sample. During our group’s analysis of the samples, we investigated various parts of 3 different specimens: a controlled fly, a mutated fly with Hippo 1, and a mutated fly with Hippo 2. Our primary focus when photographing the flies was the antenna, eyes, abdomen, upper body, legs and wings.
Meg: For me, I have found this experience to be an insightful one. I’ve learnt how to use new equipment and how to perform different experiments that I never would have done before. A highlight of experience here would have to be extracting the DNA from the specimen. Going on the tour through the Peter MacCallum Cancer Institute was a really insightful experience and I really enjoyed looking at the fruit flies through the microscopes and learning how to distinguish them from male and female. This GTAC experience for me has been a very rewarding one as I have made countless new friends and learnt a lot of new information.
Ash: My personal experience at GTAC has been very fascinating, educational and social, over these past 5 days I’ve been exposed to new equipment, people and information that has allowed me to develop my knowledge in fields that I had not yet been exposed to. My favourite part of the program was analysing the mutations both under the stereo and SEM microscopes and identifying the differences between them. Overall, I have thoroughly enjoyed my experience, I have made some new friends and it has helped me to get a clearer understanding of possible future career paths.
Darian: this whole experience has shown me a new side to being a scientist and I feel like being a scientist would be a good job to do.
Sam: Throughout this week, I’ve been able to experience things that I thought wouldn’t be possible for me to accomplish. I came to GTAC so I could learn more about science, as I find it to be a fun and fascinating subject. GTAC has been able to allow access to a more advanced range of equipment. I’m currently doing VCE biology, and throughout this week, I’ve learnt new things that have improved my understanding of biology as a whole. This experience has taught new skills and terms that will be beneficial for future use. Over the week I have learnt how to use a pipette and centrifuge at its fullest extent. Visiting the Peter MacCallum Cancer Institute was an outstanding experience, showing me what it’s like to work in the science industry, also showing what it takes to get into that position. Overall this experience has been one that I won’t forget.
Kiara: I view my experience at GTAC as a big privilege, being able to attend was amazing. The experience I liked the most doing was workshops in all different places as it helped me understand different aspects of science.
Ella: I really enjoyed my time at GTAC. I had fun learning about DNA and being exposed to something I wouldn’t otherwise know about, learning about mutations is pretty interesting!! The people I worked with are really nice and fun. Overall I had a great time.