Composite
Chaperons
Part:BBa_K1462370:Design
Designed by: Ye Peng & Junjie Zhang Group: iGEM14_SCUT (2014-10-10)
pTEF2+GroES+tADH1+pTEF2+GroEL+tADH1
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]
Design Notes
Functional expression of RuBisCo would be strongly stimulated in the presence of its chaperons. Compared with the endogenous chaperon couple of S.cerevisiae, Hsp60/Hsp10, existing in mitochondrial matrix, EroGL/EroGS, two E.coli protein-folding chaperones, are much more efficient since RuBisCo expression requires their activity in the cytosol.
Figure 1 . The schematic of mechanism of the molecular chaperons GroES and GroEL. Firstly, an ATP binds with GroEL and the ATP-GroEL immediately captures the other unit GroES and there comes the GroEL-GroES-ATP complex.With the help of the complex, the unfolded RuBisCo protein can then properly fold and make a difference in our project.
Source
EroGL and EroGS are two E.coli protein-folding chaperons. We got pTEF2 and tADH1 from kits, GroEL and GroES from a synthesis company.
References
Víctor Guadalupe-Medina, H Wouter Wisselink, Marijke AH Luttik, et al. : Carbon dioxide fixation by Calvin-Cycle enzymes improves ethanol yield in yeast. Biotechnology for Biofuels 2013 6:125.