Difference between revisions of "Part:BBa K1981201"
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===1. Usage and Biology===
 
===1. Usage and Biology===
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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.
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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. We firstly isolated promoter lsr from E.coli MG1655. GFP BBa_E0040 and double terminator BBa_B0015 are standard part offered by iGEM. Then we succeesflly constrcuted plsr+ GFP +double terminator by using Homologous recombination technology.  
  
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[[Image:AI-2 Response Device AConstruction Map by NKU China.png|800px|thumb|center|'''Figure 1:''' Schematic overview of the AI-2 Response Device A.]]
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[[Image:AI-2 Response Device AConstruction Map by NKU China.png|900px|thumb|center|'''Figure 1:''' Schematic overview of the AI-2 Response Device A.]]
 
===2. Characterization===
 
===2. Characterization===
 
===2.1 Construction verification===
 
===2.1 Construction verification===

Revision as of 00:49, 15 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


1. Usage and Biology

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. We firstly isolated promoter lsr from E.coli MG1655. GFP BBa_E0040 and double terminator BBa_B0015 are standard part offered by iGEM. Then we succeesflly constrcuted plsr+ GFP +double terminator by using Homologous recombination technology.

Figure 1: Schematic overview of the AI-2 Response Device A.

2. Characterization

2.1 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.


2.2 Response ability to exogenously added AI-2

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: GFP expression of AI-2 Response Device A when add exogenous AI-2.

2.3 Co-culture with AI-2 Controllers

2.3.1 Co-culture with AI-2 Consumers

AI-2 Consumers was constructed by iGEM 2016 NKU_China by overexpression the components responsible for AI-2 uptake (lsrACDB), phosphorylation (lsrK), and degradation (lsrFG), which can directly absorb and degrade AI-2 in the nature or artificial environment. When E.coli consisting of AI-2 Response Device A are co-cultured with AI-2 Consumers, the GFP expression of AI-2 Response Device A is directly decreased compared to control group.

Figure 1: GFP expression of AI-2 Response Device A when co-cultured with AI-2 Consumers.

2.3.2 Co-culture with AI-2 Suppliers

AI-2 Suppliers was constructed by iGEM 2016 NKU_China by overexpression the components responsible for AI-2 production (luxS, mtn), which can directly supply and enrich the AI-2 molecular level in the nature or artificial environment. When E.coli consisting of AI-2 Response Device A are co-cultured with AI-2 Suppliers, the GFP expression of AI-2 Response Device A is directly increased compared to control group.

Figure 1: GFP expression of AI-2 Response Device A when co-cultured with AI-2 Suppliers.