Team:Chiba/Sender experiments/Senders(XL10Gold) T9002(JW1908)
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Results
Reaction temparature:37°C
Sender culture:500μL,Receiver:500μL
- 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.
Sender culture:100μL,Receiver:1000μL
Sender culture:10μL,Receiver:1000μL
Reaction temparature:30°C
- Sender culture:500μL, Receiver culture:500μL
- Fig. 7
- Result
- The highest maximum fluorescence intensity was caused by RhlI gene,followed by LuxI(with LVA) and LasI gene.
- 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
- Maximum fluorescence intensity caused by RhlI gene was higher than that caused by LuxI gene.There are two reasons:
- RhlI was more active at this experimental condition.
- LuxI was well degraded by protease.
- We concluded that LVA-tag effective in this condition.
- Fig. 8
- 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:25°C
Sender culture:500μL,Receiver culture:500μL
- Fig.9
- results
- The value of fluoroscence intensity by LuxI, RhlI, LasI gene at 25°c were almost the same as negative control at 30 and 37°c.
- The last value of fluoroscence intensity by LasI gene was a half of LuxI and RhlI.
- Discussion
- Low temperature of 25°c caused decrease of production efficiency of AHL synthase.
- Static culture keep sender and receiver from shaking, in the end , expression of GFP decreased.
- Fig.10
- Result
- The fluoroscence intensity by RhlI with LVAtag is less than RhlI without LVAtag.
- Discussion
- LVAtag is effective in this condition.