Part:BBa_K2271103

pex15

Biology and Usage

This part contains a truncated version of the peroxisomal membrane protein pex15 (aa315-383).

P: This part is a truncated version of the S. cerevisiae Pex15 which is a membrane peroxisomal membrane Protein. This version could be used as a membrane anchor for applications at the peroxisomal membrane. Both C- and N-Terminus could be used for a fusion. The N-Terminus is located in the cytoplasm and the C-Terminus in the peroxisomal lumen.

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). Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]