Team:LambertGA/Parts
Our 2016 iGEM team used existing Biobrick parts from the Uppsala team (BBa_K1033906) and also the Endy Lab (BBa_M0050-LAA, M0052-DAS). We were trying to further Edinburgh’s work by expanding upon their experiments with GFP and the degradation tags. We sought to do this by replacing GFP with the tsPurple chromoprotein and characterizing the ClpXP system. We attempted to characterize these previous parts but ran into some obstacles while working with them.
We first obtained the tsPurple sequence from the 2013 Uppsala iGEM wiki page. We ordered the sequence online from IDT, hydrated it, and proceeded to carry out the 3A assembly procedures to create our constructs. We had the tsPurple with no degradation tag, tsPurple with the LAA degradation tag, and tsPurple with the DAS degradation tag. The degradation tags were obtained from the Endy Lab sequencing and correspond to strong degradation for LAA and moderate for DAS.
We sought to improve upon the LAA and DAS degradation tags by showing their relative strength in degrading the tsPurple chromoprotein and we also sought to improve upon the characterization of the tsPurple gene so that its relative degradation may be present for anyone who needs it.
We first obtained the tsPurple sequence from the 2013 Uppsala iGEM wiki page. We ordered the sequence online from IDT, hydrated it, and proceeded to carry out the 3A assembly procedures to create our constructs. We had the tsPurple with no degradation tag, tsPurple with the LAA degradation tag, and tsPurple with the DAS degradation tag. The degradation tags were obtained from the Endy Lab sequencing and correspond to strong degradation for LAA and moderate for DAS.
We sought to improve upon the LAA and DAS degradation tags by showing their relative strength in degrading the tsPurple chromoprotein and we also sought to improve upon the characterization of the tsPurple gene so that its relative degradation may be present for anyone who needs it.
