Difference between revisions of "Part:BBa K2485012"
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===Construct Design=== | ===Construct Design=== | ||
The Voltage protein we designed is a triple fusion protein. Its schematic is shown in Figure 1. Each domain has a distinct function. From the amino terminus to the carboxy terminus, these domains are the proteorhodopsin domain, the cPT domain, and the RFP domain. The first domain contains a green-light absorbing proteorhodopsin. We chose to use a proteorhodopsin rather than a channelrhodopsin or a bacteriorhodopsin, because eukaryotic channelrhodopsins are difficult to express in E. Coli due to the requirement of post-translational modifications, and because bacteriorhodopsins form a hexagonal lattice structure which is very important to functionality, less so than the trimeric structure formed by proteorhodopsin.1,2 <br /> <br /> | The Voltage protein we designed is a triple fusion protein. Its schematic is shown in Figure 1. Each domain has a distinct function. From the amino terminus to the carboxy terminus, these domains are the proteorhodopsin domain, the cPT domain, and the RFP domain. The first domain contains a green-light absorbing proteorhodopsin. We chose to use a proteorhodopsin rather than a channelrhodopsin or a bacteriorhodopsin, because eukaryotic channelrhodopsins are difficult to express in E. Coli due to the requirement of post-translational modifications, and because bacteriorhodopsins form a hexagonal lattice structure which is very important to functionality, less so than the trimeric structure formed by proteorhodopsin.1,2 <br /> <br /> | ||
Revision as of 04:59, 27 October 2017
Proteorhodopsin-cPT-RFP
This is a triple fusion protein containing green light absorbing proteorhodopsin, the cPT region of IcsA, and Fresno RFP. It is intended to function as a unipolarly expressing proton pump. The cPT region directs the localization of the protein to the old pole of E. Coli while the proteorhodopsin pumps protons out of the cell in response to green light (must be grown in 10uM all-trans retinal). RFP has also been tagged for fluorescent visualization. We also have referred to this as the Voltage protein.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1117
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]