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



Sender culture：500&mu;L,Receiver：500&mu;L







 * 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
 * 1) Maximum fluorescence intensity was vary,however,time before gfp expression doesn't differ.
 * 2) We concluded that LasI gene(BBa_K084007) was not work well at this condition.

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.
 * Fig. 2
 * Results
 * Discussion
 * The rate of AHL synthesis decreased by the degradation of autoinducer synthase,however,the length of time before gfp expression is same.

Cultures containing cells transformed with RhlI gene(BBa_K084008) and RhlI+LVA(BBa_K084009) draw the same transfer curve.
 * Fig. 3
 * Results
 * Discussion
 * We concluded that there was no effect of LVA-tag on time before gfp expression.



Reaction temparature:30°C

 * Sender culture：500&mu;L, Receiver culture：500&mu;L


 * Fig. 7
 * 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. 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.

Sender culture：500μL,Receiver culture：500μL

 * Fig.9
 * results
 * 1) The value of fluoroscence intensity by LuxI, RhlI, LasI gene at 25&deg;c were almost the same as negative control at 30 and 37&deg;c.
 * 2) The last value of fluoroscence intensity by LasI gene was a half of LuxI and RhlI.
 * Discussion
 * 1) Low temperature of 25&deg;c caused decrease of production efficiency of AHL synthase.
 * 2) 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.

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