Difference between revisions of "Part:BBa K2271103"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K2271103 short</partinfo> | <partinfo>BBa_K2271103 short</partinfo> | ||
| + | ===Biology and Usage=== | ||
| + | This part contains a truncated version of the peroxisomal membrane protein pex15 (aa315-383). | ||
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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. | 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. | 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. | ||
| − | + | <br> | |
| + | Additionaly we tested this part as a membrane anchor for the v-SNARE [https://parts.igem.org/Part:BBa_K2271060 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). | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
Revision as of 23:58, 29 October 2017
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]