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The location of the Fun26 transporter in H. polymorpha

Kreeft, Jorn (2024) The location of the Fun26 transporter in H. polymorpha. Bachelor's Research Project (period 2b), Life Science and Technology.


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In this research bachelor project, we investigated if the Fun26 transporter is also located in the peroxisomal membrane. Originally is the Fun26 transporter a vacuolar membrane transporter. A mass spectrometry study was done on protein transcription levels and the abundance of proteins in a WT baker’s yeast strain and a pex3 baker’s yeast strain. The protein transcription levels and protein abundance levels are compared to each other for both strains. The pex3 strain has no functional peroxisomes present in the cells and has a reduction in the protein abundance level of the Fun26 transporter compared to the WT strain whereas the transcription levels of the Fun26 protein were roughly the same for both strains. Fun26 is a transporter of nucleosides and nucleobases which are used in peroxisomes as substrate by peroxisomal enzymes. The MEGFP-FUN26 gene (N-terminal fusion) and the FUN26-MEGFP gene (C-terminal fusion) were transformed into the genome of H. polymorpha ku80 PEX3-MKATE2. Fluorescence microscopy was done to colocalize the meGFP fusion proteins with Fun26 and Pex3-mKate2 (if this is the case). Western blot was also done to see if the meGFP fusion proteins with Fun26 are present in the strains. There is no colocalization between meGFP and mKate2 so the Fun26 transporter is not localized in the peroxisomal membrane. Observation of meGFP fusion proteins with Fun26 is really difficult to prove and therefore it is difficult to draw conclusions on the location of the Fun26 tr

Item Type: Thesis (Bachelor's Research Project (period 2b))
Supervisor name: Kiel, J.A.K.W. and Klei, I.J. van der and Boer, R. de
Degree programme: Life Science and Technology
Thesis type: Bachelor's Research Project (period 2b)
Language: English
Date Deposited: 04 Jun 2024 09:32
Last Modified: 04 Jun 2024 09:32

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