Difference between revisions of "Part:BBa T9002"
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<partinfo>BBa_T9002 parameters</partinfo>
 
<partinfo>BBa_T9002 parameters</partinfo>
 
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__NOTOC__
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<partinfo>BBa_T9002 short</partinfo>
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Characterized by Peking_2019
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Experiment procedure:
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1. E. coli DH5α transferred with PSB1C3-T9002 were incubated in LB liquid medium at 37℃, 220rpm overnight
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2. Grown culture was 100-fold diluted in M9 medium for the same condition for 3 hour.
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3. Culture was 40-fold diluted in M9 medium containing different concentration of AHL,
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4. 200 µl aliquots of each of the cultures were transferred into a flat-bottomed 96 well plate
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5. FI and OD600 was measured for 10 hours in microplate reader.
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https://2019.igem.org/wiki/images/c/cb/T--Peking--QS2.png
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Figure1 The
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We used part T9002 to characterizing quorum sensing system. But we found that this part has an obvious homologous recombination. So we changed the terminator of GFP to avoid it.
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https://2019.igem.org/wiki/images/e/e2/T--Peking--QS.png
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Figure2 The design of quorum sensing system
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https://2019.igem.org/wiki/images/1/10/T--Peking--QS1.png
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Figure3 We use this part of a AHL receiver. With the distance between donor and receiver cells, the GFP expression level is significantly changed.
 +
Experiment procedure:
 +
1. Incubate donor and receiver cells in LB medium overnight.
 +
2. Grown culture was 100-fold diluted in M9 medium.
 +
3. 5 μL donor cell was dropped in the LB solid medium.
 +
4. 5 Μl receptor cell was dropped in different distances in the same LB solid medium

Revision as of 21:57, 20 October 2019


GFP Producer Controlled by 3OC6HSL Receiver Device

The luxR based receiver, F2620 (formerly I13270), controls the production of GFP. The GFP protein generator is the same as that found in I7101.

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 1004
    Illegal BsaI.rc site found at 1732



GFP Producer Controlled by 3OC6HSL Receiver Device Characterized by Peking_2019

Experiment procedure: 1. E. coli DH5α transferred with PSB1C3-T9002 were incubated in LB liquid medium at 37℃, 220rpm overnight 2. Grown culture was 100-fold diluted in M9 medium for the same condition for 3 hour. 3. Culture was 40-fold diluted in M9 medium containing different concentration of AHL, 4. 200 µl aliquots of each of the cultures were transferred into a flat-bottomed 96 well plate 5. FI and OD600 was measured for 10 hours in microplate reader. T--Peking--QS2.png Figure1 The We used part T9002 to characterizing quorum sensing system. But we found that this part has an obvious homologous recombination. So we changed the terminator of GFP to avoid it. T--Peking--QS.png Figure2 The design of quorum sensing system T--Peking--QS1.png Figure3 We use this part of a AHL receiver. With the distance between donor and receiver cells, the GFP expression level is significantly changed. Experiment procedure: 1. Incubate donor and receiver cells in LB medium overnight. 2. Grown culture was 100-fold diluted in M9 medium. 3. 5 μL donor cell was dropped in the LB solid medium. 4. 5 Μl receptor cell was dropped in different distances in the same LB solid medium