GFP purification

1. GFP purification Charles Latrobe College Mariam,Sara,Hoda,Aisha,Kira and Alana.

2. What we believed science was These are Quotes from each student about what each thought: “science is really hard” “scientists are nerdy” “scientists are intelligent” “scientists are smarter than me.”

3. How is this project related to a real world problem? When mitochondria don’t function properly it can lead to diseases like Huntington and Parkinson 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 they lead to different diseases.

4. What do we mean by integrated science? Integrated science is when you use more than one different sciences An example is crystallography, we are trying to figure out a biology question while using chemistry and physics. Researchers that are interested in biology can use chemistry and physics to learn about the human body is different ways.

5. The project goal the project goal was to purify the protein GFP. We want to purify proteins so that we can crystalize them to determine there structure. We need to know the structure to see how the amino acids are important and how the protein works.

6. GFP • Green Fluorescent Protein (GFP) has existed for more than one hundred and sixty million years in one species of jellyfish, Aequorea victoria. The protein is found in the photo organs of Aequorea 24/08/2014

7. What is GFP used for? • GFP is used as a marker to tag proteins so you can see them better. 24/08/2014

8. GFP Purification Protocol • Resuspend cell pellet • Add the aliquot of lysozyme • Incubate for 10mins at room temperature • We added DNase • Incubate for 10 minutes at room temperature • Sonicate lysate • We separated the soluble fraction and purified the protein 24/08/2014

9. GFP Purification Protocol • Bind the protein to the Ni-column • Wash off all other proteins • Then we got back the GFP from the column 24/08/2014

10. GFP Purification GFP 24/08/2014

11. Microscopic cells • You use the blue UV light so you can produce the fluorescent colour. • The microscope we used was called spinning disc microscope. • You can change the colour of the UV light if that particular colour is in that cell. • The green light from the microscope is less energetic then the blue light and that is what makes the cell green. • The red light from the microscope is less energetic than the yellow light and that’s what turns the mitochondria red. 24/08/2014

12. mitochondria • We looked at the mitochondria in cells. • We saw punta on the mitochondria. • We saw that the mitochondria we very dynamic. • We saw that the mitochondria moved a lot. • If the mitochondria don’t move a lot and are small, it means the cells are sick. • The mitochondria is normally around the nucleus. 24/08/2014

13. Further Areas or Questions to Investigate From the results we gained, we can see that there is a need in research for the following: How can we see proteins better? How can we explore GFP in other ways? Are there other proteins that are found at the mitochondria when they divide?

14. What was interesting about the science that is happening in this lab? • The most interesting part in the science lab was using all the different types of equipment and the types of buffers. • We also looked at the gel under the UV light and saw a fluorescent GFP band • Viviane showed us the crystalised proteins under the microscope • `

15. Why Should We care about the Research? We should care about research since it plays a big part in our everyday life. We need to know more about how the human body works. We need to know more about how mitochondria work in a cell to keep the energy supply How mitochondria move in a cell is really important and when this doesn’t work properly, diseases can occur

16. How is it relevant to Community and Society? It brings more knowledge to our community We want to know how proteins function in diseases We want to know how the proteins in mitochondria affect Huntington disease and Parkinson disease

17. How Has Our View of Science or Scientists Changed? 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.

18. What Have We Learnt? Measures and read data accurately Team Work Problem Solving Observation

19. What choices have opened up to me In Possible Career Choices? We have seen how scientists work in the labs and we’re interested in doing more science We know that you can do science after school And we know that there are several different careers in science that allow you to work with different types of equipment

20. How has being part of GTP changed my appreciation for physical science? We now appreciate science a whole lot more because we understand what they do for the world, and how hard they work in the process, for e.g.. PhD students who discover new things to make a difference.

21. Acknowledgements A special thanks to Viviane Richter our mentor for guiding us in the laboratory and helping us understand the process of purifying proteins Jill, our career pathways teacher for guiding us and helping us get here Eroia, for organizing this experience and giving us the opportunity to explore in the lab Prof Mike Ryan, Latrobe university, LIMS