Difference between revisions of "Part:BBa K2271103"

Revision as of 18:26, 30 October 2017

pex15

Biology and Usage

Figure 1. The micrscopy showing a localisation of the fluorescence protein Venus with the typical peroxisomal shape. This indicates, an peroxisomal membrane licalisation of Venus-Pex15

This part contains a truncated version of Pex15 (315-383) from S. Cerevisiea . Halbach et al. (2006)
Pex15 is a tail anchord peroxisomal membrane protein with its c-terminus directed to the peroxisomal lumen and its n-terminus directed to the cytosol. It is required for peroxisome biogenesis. In cells lacking Pex15 peroxisomal matrix proteins mislocalize to the cytosol while overexpression results in impaired peroxisome assembly. Elgersma Y, et al. (1997)

Experimental design

The functionality of this part was validated by microscopy. For the microscopy mVenus was fused to the C-Terminus of Pex15. We could observe a peroxisomal typical localisation in the cell.
Additionaly we tested this part as a membrane anchor for the v-SNARE Snc1. We co-expressed the Snc1-Pex15 construct with GUS-PTS1 to perform a GUS Assay. The secreted GUS in the supernatant was measured with the turnover of 4-methylumbelliferyl-beta-D-glucuronide to 4-methyl umbelliferone (4-MU). The fluorescent 4-MU was measured with a plate reader (excitation: 365 nm, emission: 465 nm).

Figure 2. The micrscopy showing an localisation of the fluorescence protein mVenus with the typical peroxisomal shape. This indicates, an peroxisomal membrane licalisation of mVenus-Pex15

Sequence and Features