Difference between revisions of "Part:BBa K4779005"

Latest revision as of 19:36, 11 October 2023

UAScit-prm1

UAScit-prm1 is a new synthetic hybrid promoter that improves the basal activity and induced expression level of the prm1 promoter. It consists of two tandem modules, the enhancer element UAScit and the prm1 promoter, where UAScit is a constitutive promoter. The fluorescence intensity of the engineered yeast strain BY4741-pRS415-UAScit-prm1-GFP-CYC1 was assessed using flow cytometry, and the data were analyzed with FlowJo and GraphPad Prism software. The findings revealed a 13.2% increase in basal activity and a 76.9% increase in induced expression levels compared to the prm1 promoter. This demonstrates that the constitutive enhancer element UAScit enhances both basal and induced expression levels, regardless of whether it is placed in front of inducible or constitutive promoters.

Characterization

The upstream activation element UAScit was concatenated in frame with prm1 promoter, named UAScit-pprm1. UAScit, an identified enhancer element, is a 275bp sequence upstream of the CIT1 start codon. Then the activity of these promoters can be quantitatively measured through flow cytometer. The results are shown in Figure 5-1.For UAScit, although the pheromone-induced synthesis promoter UAScit-prm1 will increase the background expression level to a certain extent, it can greatly increase its induced expression level, compared with its corresponding core promoter prm1 basic activity increased by 20.3%, and the induced expression level increased by 27.4%.

Sequence and Features