Difference between revisions of "Part:BBa K1981201"

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===Characterization===
 
===Characterization===
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===Construction verification===
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AI-2 Response Device consists of the AI-2 quorum sensor-inducible promoter BBa_K1981101(249), a GFP coding sequence BBa_E0040(747bp), a double terminator BBa_B0015(115). The total length of AI-2 Response Device A is 1111bp.
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[[Image:NKU China Device PCR Colony Verification.png|800px|thumb|center|'''Figure 1:''' Colony PCR Verification for AI-2 Response Device A.]]
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===Construction verification===
 
===Construction verification===
  
 
[[Image:AI-2 Response Device B Construction Map by NKU China.png|800px|thumb|center|'''Figure 10:''' Activity assay of each purified fraction of the cultivation with ECOL. Samples were re-buffered into H<sub>2</sub>O and the protein amount in each fraction has been adjusted. The measurements were done using the [http://2012.igem.org/Team:Bielefeld-Germany/Protocols/Analytics#General_setup_of_enzyme_activity_measurements/ standard activity assay protocol] over night. The first number indicates the percentage of used elution buffer, whereas the second number stands for the fraction number of this elution.]]
 
[[Image:AI-2 Response Device B Construction Map by NKU China.png|800px|thumb|center|'''Figure 10:''' Activity assay of each purified fraction of the cultivation with ECOL. Samples were re-buffered into H<sub>2</sub>O and the protein amount in each fraction has been adjusted. The measurements were done using the [http://2012.igem.org/Team:Bielefeld-Germany/Protocols/Analytics#General_setup_of_enzyme_activity_measurements/ standard activity assay protocol] over night. The first number indicates the percentage of used elution buffer, whereas the second number stands for the fraction number of this elution.]]

Revision as of 22:55, 14 October 2016


Autoinducer-2 Response Device A

This composite part consists of the AI-2 (autoinducer-2) quorum sensor-inducible promoter BBa_K1981101, a GFP coding sequence BBa_E0040, a double terminator BBa_B0015. In AI-2 Response Device A, GFP expression is under the control of promoter, plsr. When phospho-AI-2 binds LsrR, expression of GFP ensues. The expression of GFP can directly response to the AI-2 level in the environment, which is an alternative way to reflect the AI-2 concentration in the nature or artificial environment.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 898


Usage and Biology

This device is used to reflect AI-2 concentration in the nature or artificial environment. By co-culturing ‘AI-2 controllers’ with E.coli consisting of AI-2 Response Device A , we can know how the ‘controller cells’ manipulate the AI-2 level in the extracellular environment by simply measure the GFP expression level.

Characterization

Construction verification

AI-2 Response Device consists of the AI-2 quorum sensor-inducible promoter BBa_K1981101(249), a GFP coding sequence BBa_E0040(747bp), a double terminator BBa_B0015(115). The total length of AI-2 Response Device A is 1111bp.

Figure 1: Colony PCR Verification for AI-2 Response Device A.

Construction verification

Figure 10: Activity assay of each purified fraction of the cultivation with ECOL. Samples were re-buffered into H2O and the protein amount in each fraction has been adjusted. The measurements were done using the [http://2012.igem.org/Team:Bielefeld-Germany/Protocols/Analytics#General_setup_of_enzyme_activity_measurements/ standard activity assay protocol] over night. The first number indicates the percentage of used elution buffer, whereas the second number stands for the fraction number of this elution.