Template:NAU-CHINA/promoter
Promoter Collection
Establish expression elements in
Saccharomyces cerevisiae
Precise regulation of gene expression is pivotal to engineer a metabolic pathway, in which the promoter serves as an important regulatory element. Recent development in synthetic biology allows assembly of large DNA fragments to become much more convenient. In addition, methods on modular design and construction of biological parts were invented and widely applied during the process of constructing a metabolic pathway. These techniques make it more convenient to optimize the regulatory network of metabolic pathways.
However,The common eukaryotic model yeast Saccharomyces cerevisiae is still transcriptionally limited compared with the tools available in common bacterial hosts such as Escherichia coli.
Therefore, we constructed a lot of promoters in S. cerevisiae as standard parts, as well as a library of reporter plasmids and profiled the promoters based on the fluorescence intensity to modulating promoter activity.
- Part Name
- Short Description
- BBa_K2365031
- TDH1 promoter
- BBa_K2365033
- PDC1 promoter
- BBa_K2365039
- ADH1 promoter
- BBa_K2365040
- PGK1 promoter
- BBa_K2365036
- 2HXT7 promoter
- BBa_K2365041
- TDH3 promoter
- BBa_K2365042
- TEF2 promoter
- BBa_K2365032
- TEF1 promoter
- BBa_K2365051
- TPI1 promoter
- BBa_K2365514
- PGL1 promoter
- BBa_K2365515
- Mutant of TEF1 promoter
- BBa_K2365516
- HXT7 promoter
Figure 1. The fluorescence of the five test devices and control transformed into Escherichia coli and inoculated in LB broth was measured after 6h.
Figure 2.Constructing the promoter and GFP(s657) in the expression vector pRS423.Trough the excitation light 440-550nm to assay the fluorescence intensity. TEF1,transcriptional elongation factor promoter, TDH1,Glyceraldehyde-3-phosphate dehydrogenase promoter. PDC1,pyruvate decarboxylase promoter. TPI1,triosephosphate isomerase promoter. Mutant TEF1,transcriptional elongation factor promoter which has mutation sequence, PGL1,polygalacturonase promoter. HXT7,hexose transporter promoter, TDH3,strong Glyceraldehyde-3-phosphate dehydrogenase promoter. PGK1,phosphoglycerate kinase promoter. 2HXT7,from the Hexose transporter family,a strong promoter..
Moreover, we found an interesting phonomenon that a bottle of E. coli also expressed green fluorescence.So we transform the shuttle plasmid pRS423 into the E.coli BL21(DE3),and cultured for some time to measure the green fluorescence at 475-530nm. Through the results,We speculated that some constitutive promoters in the eukaryotic and prokaryotic system exists in the orthogonality,and we can have a future study in this topic.
Figure 3. The fluorescence of Escherichia coli BL21 which cultured after 12h.
Nevertheless, the attractiveness of yeast as a platform organism (especially for metabolic engineering applications) needs continued efforts to expand the set of tools available.
