Subcellular localization of Mtm1 in Hansenula Polymorpha

Kruijer, Eva (2024) Subcellular localization of Mtm1 in Hansenula Polymorpha. Bachelor's Research Project (period 2b), Life Science and Technology.

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Abstract

Peroxisomes are organelles found in eukaryotic cells. Transporter proteins are needed to transport solutes across the peroxisomal membrane. There is still a lot unknown about the metabolite transport. In this research the localization of the pyridoxal 5’phosphate transporter protein Mtm1 was studied using fluorescence microscopy. To study this plasmids encoding for a N-terminal and encoding for a C-terminal fusion of Mtm1 to meGFP was made. This plasmid was transformed into Hansenula polymorpha Δku80 with a peroxisomal marker Pex3-mKate2. These transformants were then used for a western blot and fluorescence microscopy. The western blot was detected using and AP-conjugate. The cells for fluorescence microscopy were grown on glucose, methanol and on glucose after which they were stained with MitoTracker Red. The observed location of the GFP was not colocalized with the mKate2 location. The western blot showed the presence of the fusion protein for both the N- and C-terminal fusion. From the obtained data it was concluded that there was no colocalization between the fusion protein and the peroxisomes. It was also found that the Mtm1 is located at the mitochondria for the cells grown on glucose.

Item Type:	Thesis (Bachelor's Research Project (period 2b))
Supervisor name:	Klei, I.J. van der and Kiel, J.A.K.W.
Degree programme:	Life Science and Technology
Thesis type:	Bachelor's Research Project (period 2b)
Language:	English
Date Deposited:	04 Jun 2024 10:31
Last Modified:	04 Jun 2024 10:31
URI:	https://fse.studenttheses.ub.rug.nl/id/eprint/32473

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