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

<html><link rel="stylesheet" href="https://2008.igem.org/wiki/index.php?title=User:Hiroki/style.css&action=raw&ctype=text/css" type="text/css" /></html>
[[Image:Chiba-U.gif]]

{| style="color:white;background-color:Maroon" cellpadding="3" cellspacing="3" border="1" bordercolor="white" width="100%" align="center"
!align="center"|[[Team:Chiba|Home]]
!align="center"|[[Team:Chiba/Team|The Team]]
!align="center"|[[Team:Chiba/Project|The Project]]
!align="center"|[[Team:Chiba/Parts|Parts Submitted to the Registry]]
!align="center"|[[Team:Chiba/Reference|Reference]]
!align="center"|[[Team:Chiba/Notebook|Notebook]]
!align="center"|[[Team:Chiba/Acknowledgements|Acknowledgements]]
|}

__NOTOC__
==Results==
===Reaction temparature:37°C===
====Sender culture：500&mu;L,Receiver：500&mu;L====
[[Image:Chiba_communication_XL10Gol_37_RS1_01.gif‎|thumb|left|'''Fig.1'''　　<br>E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,37&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]

[[Image:Chiba_talks_XL10G_37_RS1_02.gif‎|thumb|left|'''Fig.2'''　　<br>E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,37&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]

[[Image:Chiba_talks_XL10G_37_RS1_03.gif‎|thumb|left|'''Fig.3'''　　<br>E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,37&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
<br clear="all">

*Fig. 1
*results
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.
*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
*Results
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.
*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([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

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


<br clear=all>
[[Image:Chiba_talks_XL10Gold_37_RS1_0917_01.gif|thumb|left|'''Fig.4'''　　<br>E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,37°C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
[[Image:Chiba_talks_XL10Gold_37_RS1_0917_02.gif|thumb|left|'''Fig.5'''　　<br>E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,37°C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]


<br clear=all>

====Sender culture：100&mu;L,Receiver：1000&mu;L====
[[Image:Chiba_talks_XL10Gold_37_RS1_01.gif|thumb|left|'''Fig. 6'''　　
<br>E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,37&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
<br clear=all>

====Sender culture：10&mu;L,Receiver：1000&mu;L====
===Reaction temparature:30°C===
*Sender culture：500&mu;L, Receiver culture：500&mu;L
[[Image:Chiba_talks_XL10G_30-1_RS1_01.gif‎‎|thumb|left|'''Fig.7'''　　<br>E.coli strain,BBa_K084007:XL10Gold,BBa_T9002:JW1908,30°C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
[[Image:Chiba_talks_XL10G_30-1_RS1_02.gif‎‎‎|thumb|left|'''Fig.8'''　　<br>E.coli strain,BBa_K084007:XL10Gold,BBa_T9002:JW1908,30°C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
<br clear="all">

*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&deg;C===
====Sender culture：500μL,Receiver culture：500μL====
[[Image:Chiba_talks_XL10Gold_25_RS1_01.gif|thumb|left|'''Fig.9'''  <br>E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,25&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
[[Image:Chiba_talks_XL10Gold_25_RS1_02.gif|thumb|left|'''Fig.10''' <br>　E.coli strain,Senders:XL10Gold,BBa_T9002:JW1908,25&ded;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
<br clear=all>
*Fig.9
**result
***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. 
***The last value of fluoroscence intensity by LasI gene was a half of LuxI and RhlI.
**Discussion
***Low temperature of 25&deg;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.

<br clear=all>
[[Team:Chiba/Project/Experiments:Sender_Crosstalk||Back to Sender experiment and result]]