Part:BBa_K1462320:Design
pGAL1+PRK+GBD-ligand+tADH1
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 649
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 150
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
A phosphoribulokinase is an enzyme that catalyzes the chemical reaction ATP + D-ribulose 5-phosphate → ADP + D-ribulose 1,5-bisphosphate This enzyme belongs to the family of transferases, specifically those transferring phosphorus-containing groups (phosphotransferases) with an alcohol group as acceptor. This enzyme participates in carbon fixation. As is stated in previous research, increased expression levels of PKR in yeast result in an overall positive effect on the ethanol yield. In the meantime, though, these would bring a small metabolic burden to the host cell. Therefore, its expression level should be controlled. That is why we utilize a inducible promoter to construct the recombinant.
Source
The PRK is from Spinacia oleracea, catalyting the first reaction of the pathway in outer mitochondrial membrane of our project.
We got pGAL1 and tADH1 from kits, PRK from a synthesis company and, GBD ligand by overlap PCR.