Difference between revisions of "Part:BBa K553001"
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==Improvement and Characterization ==
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Group: <b>Tokyo Tech 2017</b>
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Author: Takuma Yasue
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===Summary of Improvement and Characterization===
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We found that the wild type TraI(<partinfo>BBa_K553001</partinfo>)  did not suit for our final circuits because enzyme activity was weak for our project. Thus, we constructed the improved TraI(<partinfo>BBa_K2505033</partinfo>) appropriate to our final circuits. Our purpose is to enhance the activity of TraI for our final genetic circuits.
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===Result ===
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[[File:TraImutationResults.jpg|thumb|left|300px| '''Figure 1:''' '''3OC8HSL production of TraI wild type and mutant'''<br style="clear: both" />Sender <i>E. coli</i> were grown at 37℃ in liquid LB medium with 1μM of SAM. <i>E. coli</i> introduced empty vector was used as Negative Control.]]<br>
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The result of C8 production using the wild type TraI and mutants is shown in Figure 6.
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The RFU value of the TraI (K34G)-expressing cells was approximately 3-fold higher than the TraI-expressing cells. Other mutant didn’t show improvement of 3OC8HSL production. When these RFU values were converted to 3OC8HSL concentrations using the calibration curve obtained in the reagent assay (see the TraI Assay), they were calculated as 28 nM and 42 nM, respectively.
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For more information, visit our page[]
  
  

Revision as of 06:31, 1 November 2017

A. tumefaciens TraI - OC8 HLA synthase


The Agrobacterium tumefaciens TraI synthase generates the OC8 HLA.

The gene is located on the Ti plasmid together with TraR (BBa_K553000), the other regulatory component of A. Tumefaciens quorum sensing.

Download Datasheet


Improvement and Characterization

Group: Tokyo Tech 2017

Author: Takuma Yasue

Summary of Improvement and Characterization

We found that the wild type TraI(BBa_K553001) did not suit for our final circuits because enzyme activity was weak for our project. Thus, we constructed the improved TraI(BBa_K2505033) appropriate to our final circuits. Our purpose is to enhance the activity of TraI for our final genetic circuits.

Result

Figure 1: 3OC8HSL production of TraI wild type and mutant
Sender E. coli were grown at 37℃ in liquid LB medium with 1μM of SAM. E. coli introduced empty vector was used as Negative Control.

The result of C8 production using the wild type TraI and mutants is shown in Figure 6. The RFU value of the TraI (K34G)-expressing cells was approximately 3-fold higher than the TraI-expressing cells. Other mutant didn’t show improvement of 3OC8HSL production. When these RFU values were converted to 3OC8HSL concentrations using the calibration curve obtained in the reagent assay (see the TraI Assay), they were calculated as 28 nM and 42 nM, respectively. For more information, visit our page[]


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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 385
  • 1000
    COMPATIBLE WITH RFC[1000]