Table 1 shows the parts that were submitted. All biobrick parts work in S. cerevisiae and are compatible with the RFC standard.
|BBa_K2329000||Regulation||Cre-lox recombination with lox66 and lox71||Alex Hedin||473 bp|
|BBa_K2329001||RNA||gRNA, deletion in STE2||Alex Hedin||388 bp|
|BBa_K2329002||RNA||gRNA, deletion in STE3||Alex Hedin||388 bp|
Improvement of a previous biobrick part
BBa_K2329000 is an improvement of the earlier biobrick part, BBa_K740000. The improvement of the cre-lox recombinase system was done by changing the loxP site from reversible switch to a non-reversible switch. The loxP sites flanking the promoter was changed to mutated loxP sites, see Figure 1 . The new mutant loxP sites (lox66 and lox71) have showed great success since the direction of the promoter did not flip back.
Central parts which worked as expected
BBa_K2329001 and BBa_K2329002 are two central parts of our project, as replacing the native GPCRs STE2 and STE3 is vital to ensure that the only GPCR present on the yeast cell is the heterologous GPCR introduced by us.
Non submitted parts
Table 2 shows the Basic Parts which were not submitted due to time constraints. We chose to present them here anyway to perhaps help studies for iGEM teams in the future.
|BBa_K2329003||Coding||GPCR, Ri7||Alex Hedin||984 bp|
|BBa_K2329004||Coding||GPCR, Olfr1258||Alex Hedin||936 bp|
Characterization of BBa_K2329004
BBa_K2329004 was another part that got characterized. We incorporated a mouse GPCR called Olfr1258 into S. cerevisiae which showed successful results in membrane localization and response to its ligand 2-butanone.
-  Spehr M, Munger S. Olfactory receptors: G protein-coupled receptors and beyond. Journal of Neurochemistry [Internet]. 2009 [cited 10 October 2017];109(6):1570-1583. Available from: https://www.ncbi.nlm.nih.gov/pubmed/19383089