GFP purification

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GFP purification

• Charles Latrobe College
• Mariam,Sara,Hoda,Aisha,Kira
• and Alana.

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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.

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How is this project related to a real world
problem?

• When mitochondria dont 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.

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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.

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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.

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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

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What is GFP used for?

• GFP is used as a marker to tag proteins so you
  can see them better.

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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

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GFP Purification

• Protocol
• Bind the protein to the Ni-column
• Wash off all other proteins
• Then we got back the GFP from the column

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GFP Purification
GFP
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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 thats what
  turns the mitochondria red.

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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 dont move a lot and are
  small, it means the cells are sick.
• The mitochondria is normally around the nucleus.

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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?

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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

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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 doesnt work properly,
  diseases can occur

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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

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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.

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What Have We Learnt?

• Measures and read data accurately
• Team Work
• Problem Solving
• Observation

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What choices have opened up to me In Possible
Career Choices?

• We have seen how scientists work in the labs and
  were 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

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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.

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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