Team:Chalmers-Gothenburg/Parts

Chalmers Gothenburg iGEM 2017

Parts summary

Submitted parts

Table 1 shows the parts that were submitted. All biobrick parts work in S. cerevisiae and are compatible with the RFC[10] standard.

Table 1. The Basic Parts which were submitted.
Name Type Description Designer Length
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 [1]. The new mutant loxP sites (lox66 and lox71) have showed great success since the direction of the promoter did not flip back.

Figure 1. Illustrated figure of the non-reversible flip. When the double mutant loxP site is produced through recombination, the promoter will not be able to 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.

Table 2. The Basic Parts which were not submitted.
Name Type Description Designer Length
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.

References

  • [1]   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