Team:UAAAN/Composite Part
BioBrick BBa_K2998005. Characterization of the cluster of H+ producing genes, obtained from Ruminococcus albus
Considering that for the optimization of the electric power generation process in our project we require H + (to facilitate the role of the proton selective membrane) and not hydrogen in its gaseous form (H2), the strategy of removing the enzyme maturation complex gene at the end of the process. That is, creating a sequence with only the HydS and HydA2 genes, discarding HydABC.
FIG4. Scheme of the ligation process of the HydS and HydA2 genes simulated in SnapGene.
In addition, the E. coli lac promoters and the rrnB T1 terminators (frequently used as an efficient transcription terminator in several cloning vectors) and T7Te (early transcription terminator Phage T7) were added to these sequences.
FIG5. Process of adding promoters and terminators to the gene cluster
REFERENCES
Suen, G., Stevenson, D. M., Bruce, D. C., Chertkov, O., Copeland, A., Cheng, J. F., … Weimer, P. J. (2011). Complete genome of the cellulolytic ruminal bacterium Ruminococcus albus 7. Journal of Bacteriology, 193(19), 5574–5575. https://doi.org/10.1128/JB.05621-11
Zheng, Y., Kahnt, J., Kwon, I. H., Mackie, R. I., & Thauer, R. K. (2014). Hydrogen formation and its regulation in Ruminococcus albus: Involvement of an electron-bifurcating [FeFe]-hydrogenase, of a non-electron-bifurcating [FeFe]-hydrogenase, and of a putative hydrogen-sensing [FeFe]-hydrogenase. Journal of Bacteriology, 196(22), 3840–3852. https://doi.org/10.1128/JB.02070-14