Part:BBa_K2329004
The GPCR Olfr1258
This part was never submitted due to time constraints, but is characterized.
Olfactory receptor 1258 is a GPCR receptors that can interact and detect odorant molecules in a sense of smell. The olfactory receptor lays within the family G-protein-coupled receptors (GPCR), and are transmembrane located [1].
Olfr1258, olfactory receptor 1258, is a native GPCR in Mus musculus and recognizes 2-butanone. 2-Butanone is a Volatile organic compound (VOC) which means that it acts as gas in room temperature [2].
Usage and Biology
Through incorporating the GPCR, Olfr1258, into yeast, the yeast will be able to sense 2-butanone, instead of their own mating pheromones. When the butanone is sensed, the GPCR will be activated and mating between two yeast strain (a and 𝞪) will occur [3]. Check out Project description for the GPCR usage in our project.
Characterization
To replace the native GPCR and incorporate the new GPCR, Olfr1258, a CRISPR Cas9 technique was used with the gRNA presented in BBa_K2329002 page. The incorporation showed to be successful according to colony-PCR.
Membrane localization
To enable visualization of the localization of the receptor in the yeast cells, the GPCR transformed was fused to a GFP tag. The green fluorescence could then easily enable detection of the GPCR. For the yeast with Olfr1258, the membrane localization of the GPCR could be verified. In Figure 1a, it can be seen that Olfr1258 is clearly located in the outer membrane of the cell. In Figure 1b, it can be seen that these cells fluoresce in the nuclear membrane which is clearly shown when the cells in this image is dividing. This image thereby suggest that the GPCR is localized in all membranes.
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]
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