Part:BBa_K2568014
pCI/Tet+rfp
We combined rfp to promoter BBa_K1493805 to measure its strength and compared it with another 5 promoters from iGEM14_Wageningen_UR. This part is used for the characterization of the promoter BBa_K1493805.
BBa_K1493805 contains two Tet operator sites at the core and proximal position (middle and right part of the promoter) and two cI lambda operator sites at the distal site (left part of the promoter).We transduced the recombinant plasmid containing the target promoter and an rfp reporter gene into E.coli. In this experiment, we chose BioTek Synergy multifunctional enzyme mark to measure the fluorescence intensity and OD450, PBS was used as the blank control. According to the optimum detection wavelength of RFP, the parameters for detecting fluorescence intensity were: excitation wavelength of 584nm and emission wavelength of 607nm. The fluorescence / OD450 was used to measure the RFP expression level of bacteria. The results are provided in Figure 1.
Figure 1.Comparisions of the expression of 6 promoters. Expression of the promoters are ordered from high to low. Expression of the blank control is considered 0. The promoter 3 is considered unexpressed as no fluorescence expression was detected.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 685
Illegal AgeI site found at 797 - 1000COMPATIBLE WITH RFC[1000]
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