![](https://parts.igem.org/images/partbypart/icon_coding.png)
Part:BBa_K2271103
pex15
Biology and Usage
This part contains a truncated version of Pex15 (aa315-383) from S. cerevisiea . [1]
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. [2]
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).
![](/wiki/images/a/ac/Igemduscgn2017GUSassay.png)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
None |