Difference between revisions of "Part:BBa K3078100"
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SfGFP (BBa_K2541400) is used as the reporter protein in the promoter intensity test system, which allows users to replace the promoter through GoldenGate.
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This year, a promoter strength measurement device(BBa_K3078100) were created by Jilin_China. This part consists of two BbsI digestion sites, RBS and sfGFP.
 
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SfGFP (BBa_K2541400) is used as the reporter protein in the promoter intensity test system, which allows users to replace the promoter through GoldenGate.
+
Two Bbs I recongition sites on the device allow DNA to be constructted on it through GoldenGate Assmbly. And RBS and CDS of sfGFP are at the downstream of the second Bbs I site, which can be used to characterized the promotor intensity by the intensity of fluorescence.  //  Because the length of promoters in procaryotic organisms are usually short, which means these promoters are easily to synthesized in vitro. Therefore, we can easily obtain double strand DNA with two sticky ends by single stranded oligonucleotide annealing and we ligand it into the device. This method can construct short promoter measurement parts with in a large quanity and in a short time, which is suitable for the screening and characterizing of promoters. Thus, we consider that this composite part must be popular and widely used in iGEM Community.
 
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<P style="text-indent:2em;">
 
<P style="text-indent:2em;">
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SfGFP (BBa_K2541400) is used as the reporter protein in the promoter intensity test system, which allows users to replace the promoter through GoldenGate.
+
This year, we characterized the promoter P59, which haven’t be submitted to iGEM, by the help of this promoter strength measurement device. Compared with 5 promoters from Anderson Family, we identified the relative intensity of P59 to the control group, attesting this part satisified our anticipation.
 
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[[File:我他吗困死了4.png|600px|center|我他吗困死了4]]
 
[[File:我他吗困死了4.png|600px|center|我他吗困死了4]]
 
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Figure 4. BDSF inhibition of the phase transformation of C. albicans. C. albicans and the bacteria supernatant of control or Bcam0581 or BDSF standard were co-cultured in YPD medium supplementary with 10% serum in 96-well plate at 37℃ for 4 hours. After incubation, the hypha morphology was observed with inverted microscope. Three different pictures were selected from each well. A, mock control; B, bacteria supernatant of Bcam0581; C1~C3, different concentration of 0.75, 3 or 7.5 μM standard.
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Figure 1. The fluorescence change (Fluorescence/ OD600) of promoters with time. Different promoters were constructed into the same measurement device, cultured over night, diluted to OD600=0.02 and then measured the fluorescence at 528 nm with 485nm excitation wavelength.
 
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Revision as of 00:23, 22 October 2019

A promoter strength measurement intermediates.

This year, a promoter strength measurement device(BBa_K3078100) were created by Jilin_China. This part consists of two BbsI digestion sites, RBS and sfGFP.


<h5>

Two Bbs I recongition sites on the device allow DNA to be constructted on it through GoldenGate Assmbly. And RBS and CDS of sfGFP are at the downstream of the second Bbs I site, which can be used to characterized the promotor intensity by the intensity of fluorescence. // Because the length of promoters in procaryotic organisms are usually short, which means these promoters are easily to synthesized in vitro. Therefore, we can easily obtain double strand DNA with two sticky ends by single stranded oligonucleotide annealing and we ligand it into the device. This method can construct short promoter measurement parts with in a large quanity and in a short time, which is suitable for the screening and characterizing of promoters. Thus, we consider that this composite part must be popular and widely used in iGEM Community.

This year, we characterized the promoter P59, which haven’t be submitted to iGEM, by the help of this promoter strength measurement device. Compared with 5 promoters from Anderson Family, we identified the relative intensity of P59 to the control group, attesting this part satisified our anticipation.

<h5>


我他吗困死了4

Figure 1. The fluorescence change (Fluorescence/ OD600) of promoters with time. Different promoters were constructed into the same measurement device, cultured over night, diluted to OD600=0.02 and then measured the fluorescence at 528 nm with 485nm excitation wavelength.




Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 468
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]