Green Fluorescent Protein

1. Green Fluorescent Protein Mariam, Sara, Hoda, Aisha, Kira and Alana, Charles Latrobe College How is this project related to a real world problem? When mitochondria don’t function properly it can lead to Huntington’s disease and Parkinson’s disease. We use GFP to mark the proteins that are in the cells so we can see what they do and how they divide, also how they function and how different diseases are caused by proteins not working properly. Introduction In this project, we wanted to purify Green Fluorescent Protein (GFP). We can crystallise pure proteins to find the protein structure. We can find the structure by taking the crystals to the Australian Synchrotron. Finding the protein structure is important because they can show us how proteins work and how we can cure or treat different diseases. GFP is a fluorescent protein that allows us to tag proteins that we’re interested in so we can visualise them. We want to look at proteins because we want to know where they are found in the cell and how they work. In the project we were looking at mitochondria and proteins that work in division of mitochondria. How has our understanding of science and scientists changed through this experience Now that we have a lot more knowledge of what scientist do we have a better understanding of what they do and how they work in the lab, it has changed in a positive manner because we got to work in a lab surrounded by them and we realized that they work really hard to achieve their goals and make the world an healthier environment. Aim/Motivation Our aim was to: Our aim was to purify GFP What motivates scientists to investigate this area Scientists are motivated to find the structure of the proteins to find out how they function and how they are involved in disease. Diffraction Patterns of laser light through different size slits The Australian Synchrotron Ripple Tank diffraction pattern Describe what you did and how What data did you collect and what did it mean We broke open bacterial cells (lysis) and spun them down in a centrifuge. We then bound the protein to the Ni-beads which bind to a His-tag on the protein. We washed the beads and eluted the protein to get the protein pure. We ran the protein on a gel to see how pure our protein was. We stained the gel overnight so that we could see the protein. We found that there was a huge blob of protein in our purified sample and that it was pure. Data or content GFP Conclusions In conclusion, our experiment was to purify GFP and we succeeded! We could use this technique to purify other proteins in the future. Acknowledgements -Career co-ordinator Jill Dixon - The ARC Centre of Excellence for Coherent X-Ray Science: Viviane Richter, Prof Mike Ryan - The Growing Tall Poppies Program Developer: Dr Eroia Barone-Nugent Santa Maria College How has being part of GTP changed my appreciation for physical science? We now know that science is hard and it’s not all about experiments but also about the research behind it and the planning. We now know that biology and physics can come together to answer important questions. Biochemistry and Molecular Science