Difference between revisions of "Part:BBa K2995000"
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[[File:T--Waterloo--BBa K2137001-OD-corrected-fluorescence-vs-copper.png|300px|thumb|right|Figure 2: OD-corrected Fluorescence vs. Copper Concentration]] | [[File:T--Waterloo--BBa K2137001-OD-corrected-fluorescence-vs-copper.png|300px|thumb|right|Figure 2: OD-corrected Fluorescence vs. Copper Concentration]] | ||
| − | [[File:T--Waterloo--silvergraph.png|1000px|thumb| | + | [[File:T--Waterloo--silvergraph.png|1000px|thumb|left|Figure 1: OD-corrected GFP fluorescence]] |
(Excitation: 485, Emmission: 525) | (Excitation: 485, Emmission: 525) | ||
Revision as of 23:40, 21 October 2019
Paph promoter
Constitutive promoter for expression in E. coli and B. japonicum.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterization
(Waterloo iGEM 2019)
To characterize our promoter we cloned it upstream of the RBS in BBa_E0240 (RBS-GFP-Term) and compared it's GFP expression with the already well characterized BBa_J23101 promoter in BBa_I20260. To limit the impact of other variables we insured that BBa_I20260 and BBa_E0240 shared the same backbone, RBS and terminator. In addition to characterizing this promoter with respect toBBa_I20260 we also confirmed proper function of this promoter in the pRJPaph-bjGFP plasmid which was used for expression in Bradyrhizobium for our 2019 project.
Note:
empty DH5alpha was used as a negative control for GFP expression
1. Cell were grown overnight in LB media at 37 degrees
2. The following morning GFP expression was quantified using the Synergy 4 BioTek plate reader.
The graph below indicates the results from this characterization experiment:
(Excitation: 485, Emmission: 525)