Team:Chiba/Sender experiments/Senders(XL10Gold) T9002(JW1908)

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Results and Discussion

Reaction temparature:37°C

  • Sender culture：500μL,Receiver：500μL
Fig.1　　
E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,37°C
Fig.2　　
E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,37°C
Fig.3　　
E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,37°C


results

Fig. 1

Cultures containing cells transformed with LuxI gene([http://partsregistry.org/Part:BBa_K084012 BBa_K084012])and cells transformed with RhlI gene([http://partsregistry.org/Part:BBa_K084008 BBa_K084008]) express gfp and fluorescence intensity was reached to 500.Cultures containing cells transformed with LasI gene([http://partsregistry.org/Part:BBa_K084007 BBa_K084007]) weakly expressed gfp,and fluorescence intensity was not reached to a significant level.

Fig. 2

Fluorescence intensity of the culture containing cells transformed with LuxI gene([http://partsregistry.org/Part:BBa_K084012 BBa_K084012]) was lower than that of the cultures containing the cells transformed with LVA LuxI-LVA gene([http://partsregistry.org/Part:BBa_K084014 BBa_K084014]).The difference of maximum fluorescence intesity between the two cultures was 200.

Fig. 3

Cultures containing cells transformed with RhlI gene([http://partsregistry.org/Part:BBa_K084008 BBa_K084008]) and RhlI+LVA([http://partsregistry.org/Part:BBa_K084009 BBa_K084009]) draw the same transfer curve.

Discussion

Fig. 1

  • Discussion:Maximum fluorescence intensity was vary,however,time before gfp expression doesn't differ.
  • We concluded that LasI gene([http://partsregistry.org/Part:BBa_K084007 BBa_K084007]) was not work well at this condition.

Fig. 2

  • The rate of AHL synthesis decreased by the degradation of autoinducer synthase,however,the length of time before gfp expression is same.

Fig. 3

  • We concluded that there was no effect of LVA-tag on time before gfp expression.


Fig.4　　
E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,37°C
Fig.5　　
E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,37°C



Reaction temparature:30°C

  • Sender culture：500μL, Receiver culture：500μL
Fig.6　　
E.coli strain,BBa_K084007:XL10Gold,BBa_T9002:JW1908,30°C
Fig.7　　
E.coli strain,BBa_K084007:XL10Gold,BBa_T9002:JW1908,30°C


  • Fig. 6
    • Result 　
  1. The highest maximum fluorescence intensity was caused by RhlI gene,followed by LuxI(with LVA) and LasI gene.
  2. Fluorescence intensity of the two culture containing cells transformed with LuxI gene and RhlI gene　amount to 200 for 2 hour as fast as LasI gene.
    • Discussion
  1. Maximum fluorescence intensity caused by RhlI gene was higher than that caused by LuxI gene.There are two reasons:
  2. RhlI was more active at this experimental condition.
  3. LuxI was well degraded by protease.
  4. We concluded that LVA-tag effective in this condition.


  • Fig. 7
    • Result
    • Comparing RhlI with Rhl+LVAtag, fluoroscence intensity were almost same.
    • Discussion
      • In this condition, production efficiency of LuxI protein higher than of LVAtag.

Reaction temparature:25c°

Sender culture：500μL,Receiver culture：500μL

Fig.8
E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,25c°,Receiver cells/Sender cells = 1.
Fig.9
　E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,25°C,Receiver cells/Sender cells = 1.


  • Left:
    • result
      • The value of fluoroscence intensity by LuxI, RhlI, LasI gene at 25c° were almost the same as negative control at 30 and 37c°.
      • The last value of fluoroscence intensity by LasI gene was a half of LuxI and RhlI.
    • Discussion
      • Low temperature of 25c° caused decrease of production efficiency of AHL synthase.
      • またそれだけではなく静置して行ったためSenderとReceiverがよく混ざらず、その結果GFPの発現量が大幅に減少した可能性も考えられる。
  • Right:
    • Result
      • The fluoroscence intensity by RhlI with LVAtag is less than RhlI without LVAtag.
    • Discussion
      • LVAtag is effective in this condition.


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