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Master Thesis - Methods development for monitoring metal transporters in green algae Chlamydomonas reinhardtii
In this project we will adapt targeted proteomics techniques existing in our lab for the selective detection of metal transporters in extracts of metal-exposed and control Chlamydomonas reinhardtii.
Silver is a heavy metal that is highly toxic. It exerts its toxicity by inhibiting enzymes and causing oxidative stress. How silver enters a cell is not completely understood, but there is evidence that it hitchhikes copper transporter proteins (CTRs), which are essential for metal homeostasis for instance in the green algae Chlamydomonas reinhardtii.
Silver is a heavy metal that is highly toxic. It exerts its toxicity by inhibiting enzymes and causing oxidative stress. How silver enters a cell is not completely understood, but there is evidence that it hitchhikes copper transporter proteins (CTRs), which are essential for metal homeostasis for instance in the green algae Chlamydomonas reinhardtii.
Within this master thesis we will adapt existing methods for monitoring metal transporters in green algae Chlamydomonas reinhardtii. In a first step, we will select peptides specific to the cytosolic transporters CTR3 and FOX1 and optimize mass spectrometric settings for their determination. Those transporters have been seen to be upregulated in silver-exposed algae. We will then extend our investigation to membrane proteins, such as DMT1, CTR1, CTR2. This will require an optimization of the extraction procedure. If time permits we will validate our results using gene-deletion clones, lacking the respective transporter.
The candidate will learn (1) to culture green algae (2) to adapt LC-MS based targeted proteomics protocols for new target proteins and (3) to monitor specific metal transporters in metal-exposed and control green algae.
Suitable candidates for this MSc thesis project are expected to hold a BSc degree in environmental sciences or a related discipline and to have experience in laboratory work.
Within this master thesis we will adapt existing methods for monitoring metal transporters in green algae Chlamydomonas reinhardtii. In a first step, we will select peptides specific to the cytosolic transporters CTR3 and FOX1 and optimize mass spectrometric settings for their determination. Those transporters have been seen to be upregulated in silver-exposed algae. We will then extend our investigation to membrane proteins, such as DMT1, CTR1, CTR2. This will require an optimization of the extraction procedure. If time permits we will validate our results using gene-deletion clones, lacking the respective transporter.
The candidate will learn (1) to culture green algae (2) to adapt LC-MS based targeted proteomics protocols for new target proteins and (3) to monitor specific metal transporters in metal-exposed and control green algae.
Suitable candidates for this MSc thesis project are expected to hold a BSc degree in environmental sciences or a related discipline and to have experience in laboratory work.
For further information please contact Marc Suter (suter@eawag.ch) and/or visit his group page
http://www.eawag.ch/en/department/utox/organisation/bioanalytics/
This research will be performed at the department of Environmental Toxicology, Eawag, in Dübendorf.
For further information please contact Marc Suter (suter@eawag.ch) and/or visit his group page