Protein_Domain
Part:BBa_K1478000
Designed by: Fabian Frömling Group: iGEM14_Hannover (2014-09-13)
Expa4 plant secretion signal, localizes to extracellular space
Coding region for first 20 aminoacids of plant protein Exansin4. Secretion signal for plants (derived from A. thaliana). When fused to a protein, directs the protein to extracellular space.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Functional Parameters
Figure 1 shows detected fluorescence of transformed Nicotiana tabacum cells. A GFP control without protein fusion is shown in lane A-D. The signal is specific for a cytosolic protein, which is indicated by chloroplast surrounded with GFP-Signal. In contrast the plasmamembranemarker (E-H) shows a specific signal at the periphery. The signal doesnât surround the chloroplast, which shows that it is located at the cell cover. Because itâs a marker, itâs location is already known. The analyzed construct is shown in I-K. It contrast to cytoplasmic GFP, the signal appears to be in the exterior. The signal appears to diffuse between the cells, at the cell wall. This indicates that the secretion signal and the tethering at the cell wall via CBD might work.
Fig. 1: Confocal detection of fluorescence from transformed plant cells (replicate 1). Transformed cells of Nicotiana tabacum were analyzed via confocal microscopy. Column A-I: Red channel, shows chlorophyllâs autofluorescence which shows the chloroplasts. Column B-J: Green channel showing GFP. Column C-K: Red and green channel merged. Column D-L: Pseudo transmission detection. Lane A-D shows a transformed cell with raw GFP. Lane E-H shows a plasmamembranemarker (Nelson et al. 2007). Lane I-L shows the detection of the analyzed construct: Expa4-GFP-CBD. |
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Categories
Parameters
//chassis/eukaryote/athaliana
//chassis/eukaryote/ntabacum
//proteindomain/localization
//chassis/eukaryote/ntabacum
//proteindomain/localization
None |