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

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Revision as of 07:47, 30 October 2008

Results

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

• Fig. 1
• results

Cultures containing cells transformed with LuxI gene(BBa_K084012)and cells transformed with RhlI gene(BBa_K084008) express gfp and fluorescence intensity was reached to 500.Cultures containing cells transformed with LasI gene(BBa_K084007) weakly expressed gfp,and fluorescence intensity was not reached to a significant level.

• Discussion:Maximum fluorescence intensity was vary,however,time before gfp expression doesn't differ.

• We concluded that LasI gene(BBa_K084007) was not work well at this condition.

• Fig. 2
• Results

Fluorescence intensity of the culture containing cells transformed with LuxI gene(BBa_K084012) was lower than that of the cultures containing the cells transformed with LVA LuxI-LVA gene(BBa_K084014).The difference of maximum fluorescence intesity between the two cultures was 200.

• Discussion

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

• Fig. 3
• Results

Cultures containing cells transformed with RhlI gene(BBa_K084008) and RhlI+LVA(BBa_K084009) draw the same transfer curve.

• Discussion

• 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.
    • Static culture keep sender and receiver from shaking, in the end , expression of GFP decreased.

• 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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