Difference between revisions of "Part:BBa K1493804"

 
 
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<partinfo>BBa_K1493804 short</partinfo>
 
<partinfo>BBa_K1493804 short</partinfo>
  
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Promoter designed to have operators sites for the tetracycline repressor and the cI lambda repressor, based on sequences in Elowitz et. al. 2007
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This promoter contains two Tet operator sites at the distal and proximal position (left and right part of the promoter) and one cI lambda operator sites at the core site (middle part of the promoter).
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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The promoter was combined with [https://parts.igem.org/Part:BBa_I13504 GFP] to check for its functionality.
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https://static.igem.org/mediawiki/2014/f/f2/Wageningen_UR_killswitch_Pic13.jpg
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Figure 1: plates with the six diferent promters, that where created for BananaGuard, upstream of GFP. P1 is [https://parts.igem.org/Part:BBa_K1493801 BBa_K1493801], P2 is [https://parts.igem.org/Part:BBa_K1493802 BBa_K1493802], P3 is [https://parts.igem.org/Part:BBa_K1493803 BBa_K1493803], P4 is [https://parts.igem.org/Part:BBa_K1493804 BBa_K1493804], P5 is [https://parts.igem.org/Part:BBa_K1493805 BBa_K1493805], P6 is [https://parts.igem.org/Part:BBa_K1493806 BBa_K1493806].
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Figure 1 shows the P4 (promoter with CIλ and Tet operator sites) expressing GFP, suggesting that it is a functional promoter.
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This Promoter is part of a promoter set used in [http://2014.igem.org/Team:Wageningen_UR BananaGuard] listed here:
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[https://parts.igem.org/Part:BBa_K1493802 BBa_K1493801]
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[https://parts.igem.org/Part:BBa_K1493802 BBa_K1493802]
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[https://parts.igem.org/Part:BBa_K1493803 BBa_K1493803]
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[https://parts.igem.org/Part:BBa_K1493805 BBa_K1493805]
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[https://parts.igem.org/Part:BBa_K1493806 BBa_K1493806]
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===Contribution===
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Group: Team Jiangnan 2018.<br />
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Author: Yini Luo.<br />
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Summary: We combined rfp to promoter BBa_K1493804 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_K1493804.<br />
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BBa_K1493804 contains two Tet operator sites at the distal and proximal position (left and right part of the promoter) and one cI lambda operator sites at the core site (middle 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.For details, please click [https://parts.igem.org/Part:BBa_K2568013 BBa_K2568013].
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[[Image:T--Jiangnan--Promoter.png|center|thumb|400px|]]
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'''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. '''
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Latest revision as of 11:35, 13 October 2018

promoter with CIλ and Tet operator sites

Promoter designed to have operators sites for the tetracycline repressor and the cI lambda repressor, based on sequences in Elowitz et. al. 2007

This promoter contains two Tet operator sites at the distal and proximal position (left and right part of the promoter) and one cI lambda operator sites at the core site (middle part of the promoter).

Usage and Biology

The promoter was combined with GFP to check for its functionality. Wageningen_UR_killswitch_Pic13.jpg 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 P4 (promoter with CIλ and Tet operator sites) 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:

BBa_K1493801

BBa_K1493802

BBa_K1493803

BBa_K1493805

BBa_K1493806

Contribution

Group: Team Jiangnan 2018.
Author: Yini Luo.
Summary: We combined rfp to promoter BBa_K1493804 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_K1493804.
BBa_K1493804 contains two Tet operator sites at the distal and proximal position (left and right part of the promoter) and one cI lambda operator sites at the core site (middle 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.For details, please click BBa_K2568013.

T--Jiangnan--Promoter.png

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]