Difference between revisions of "Part:BBa K1493801"
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Group: Team Jiangnan 2018. | Group: Team Jiangnan 2018. | ||
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Author: Yini Luo. | Author: Yini Luo. | ||
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Summary: We combined rfp to promoter BBa_K1493801 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_K1493801. | Summary: We combined rfp to promoter BBa_K1493801 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_K1493801. | ||
BBa_K1493801 contains two TetR operator sites at the core and proximal positions (middle and right parts 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. | BBa_K1493801 contains two TetR operator sites at the core and proximal positions (middle and right parts 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. | ||
Revision as of 10:42, 7 October 2018
Promoter with Tet operator site
Promoter designed to have operators sites for the tetracycline repressor, based on sequences in Elowitz et. al. 2007
This promoter contains two TetR operator sites at the core and proximal position (middle and right part of the promoter)
Usage and Biology
The promoter was combined with GFP to check for its functionality.
Figure 1: plates with the six diferent promters, that where created for BananaGuard, upstream of GFP. P1 is BBa_K1493801, P2 is BBa_K1493802, P3 is BBa_K1493803, P4 is BBa_K1493804, P5 is BBa_K1493805, P6 is BBa_K1493806.
Figure 1 shows the P1 (Promoter with Tet operator site) expressing GFP, suggesting that it is a functional promoter.
This Promoter is part of a promoter set used in [http://2014.igem.org/Team:Wageningen_UR BananaGuard] listed here:
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]