Team:Uppsala-Sweden/Project/Data

From 2011.igem.org

(Difference between revisions)

Latest revision as of 13:52, 25 October 2011

Visit our blog:

http://igem2011uu.blogspot.com

Overview of the system

Data For Our Favorite New Parts

1. Main Page (http://partsregistry.org/wiki/index.php?title=Part:BBa_K592009) - amilCP, a blue chromoprotein, BBa_K592009. This chromoprotein from the coral Acropora millepora, amilCP, naturally exhibits strong color when expressed. The color is blue/purple and is visible to naked eye, thereby requiring no instruments to observe. The strong color is readily observed in both LB or agar culture, in less than 24 hours of incubation.

2. Main Page (http://partsregistry.org/wiki/index.php?title=Part:BBa_K592100) - Blue Fluorescent Protein (mTagBFP), BBa_K592100. This part codes for the bright blue fluorescent protein TagBFP. TagBFP is a monomeric protein with a narrow fluorescence emission spectrum with a maximum at 456 nm.

3. Main Page (http://partsregistry.org/wiki/index.php?title=Part:BBa_K592200) - Low to medium copy BioBrick standard vector, BBa_K592200. A BioBrick standard vector with low to medium copy p15A replication origin (BBa_I50032) and spectinomycin antibiotic resistance marker, usable for Lambda Red recombineering in E coli.

Data For Pre-existing Parts

1. Main page http://partsregistry.org/Part:BBa_R0011 - PLlacO – promoter, BBa_R0011: the promoter show the highest RPU-value of all characterized promoters and had the highest with IPTG.

2. Main Page (http://partsregistry.org/Part:BBa_J23113) - constitutive promoter family member, BBa_J23113 Parts J23100 through J23119 are a family of constitutive promoter parts isolated from a small combinatorial library. As expected J23113 showed a low promoter strength.

3. Main page http://partsregistry.org/Part:BBa_K592026 J23101-B0032-BFP, BBa_K592026: the constitutive promoter was used as the reference in the characterization measurement.

Characterized Parts

Our measurement with the FACS AriaII shown the following Relative Promoter Units with the Blue Fluorescent Protein (mTagBFP) as reporter, assembled with B0032 in pSB3K3-backbone transformed in TOP10 E.coli-strain.

1. Main page http://partsregistry.org/Part:BBa_K592003 - PcpcG2 promoter, BBa_K592003: measurement shows a RPU-value of 0,1.

2. Main page http://partsregistry.org/Part:BBa_K592006 - PFixK2 promoter, BBa_K592006 is activated by the blue sensor and showed no significant expression in absence of the YF1-sensor (BBa_K592004)

3. Main page http://partsregistry.org/Part:BBa_K592008 - PT5-lac Promoter, BBa_K592008: showed high expression of BFP, and showed higher expression in the presence of IPTG than without it.

@@ Line 15: / Line 15: @@
 <div class="prefix_4 grid_11 suffix_1 maincontent" id="Uppsalacontent">
+<br>
-<div class="grid_Blue_thin omega mywhite">
+<div class="grid_Green_thin omega mywhite">
-Test for light-sensor and promoter characterization
+Overview of the system
 </div>
-<br><br>
+<br>
+[[File:System overview 670.png]]
-Characterization is an important aspect of synthetic biology and it is a way of obtaining how well the expression of the system really is. Therefore we have designed two tests to perform characterization of the parts we are going to use. Both types of test are based on the expression levels of BFP. The first test is done in order to characterize the promoters that we are using in our project. The second test is done to test the output of our light sensing system. The expression levels will in both cases be measured using flow-cytometry. Here the specific details of how the test has/will been performed depending on its type.
-<div class="grid_Green_thin omega mywhite">
+<div class="grid_Red_thin omega mywhite">
-Promoter testing
+Data For Our Favorite New Parts
 </div>
-<br>
-The first test was done to see the strength of the promoters used. From the beginning it was meant to be a characterization of the promoter Pt5lac that we provide to the partsregistry. Though since the other promoters we are using which were gained from the partsregistry was not characterized yet, the decision was made that we would have to characterize all the promoters we use. Since, as said in our opinion characterization is a really important aspect of the working approach in synthetic biology.
-Every construct was made according to the approach provided by the measurement page in partsregistry besides from that the expressed protein was BFP. This means that the backbone pSB3K3, the RBS B0032 and BFP were used in every construct. The promoters tested were pT5lac, PcpcG2, PLlacO, J23113, PfixK and as reference promoter J23101. The assemblies were done according to the picture.
-The actual test was done in a somewhat different way than that presented at the partsregistry. Here we use flow-cytometry which also results in that we did not use M9 media to grow the cells but used LB instead. Also when it comes to the time of incubation of the selected colonies were 10 hours and no dilutions were made except for the settings requirements of the FACS machine.
+. Main Page (http://partsregistry.org/wiki/index.php?title=Part:BBa_K592009) - amilCP, a blue chromoprotein, BBa_K592009.
+This chromoprotein from the coral <i>Acropora millepora</i>, amilCP, naturally exhibits strong color when expressed. The color is blue/purple and is visible to naked eye, thereby requiring no instruments to observe. The strong color is readily observed in both LB or agar culture, in less than 24 hours of incubation.
+. Main Page (http://partsregistry.org/wiki/index.php?title=Part:BBa_K592100) - Blue Fluorescent Protein (mTagBFP), BBa_K592100. This part codes for the bright blue fluorescent protein TagBFP. TagBFP is a monomeric protein with a narrow fluorescence emission spectrum with a maximum at 456 nm.
+. Main Page (http://partsregistry.org/wiki/index.php?title=Part:BBa_K592200) - Low to medium copy BioBrick standard vector, BBa_K592200. A BioBrick standard vector with low to medium copy p15A replication origin (BBa_I50032) and spectinomycin antibiotic resistance marker, usable for Lambda Red recombineering in E coli.
 <div class="grid_Blue_thin omega mywhite">
-System output tests
+Data For Pre-existing Parts
 </div>
-<br>
-This test will be fully compatible with our system. The only change is that every constructed output will be done with BFP as output instead of the color output in order to get a preference of how efficient the light-sensors are. Since there are no fully functional sensor, no tests has been able to be performed. Though Both the corresponding blue and yellow output is ready to be used.
+. Main page http://partsregistry.org/Part:BBa_R0011  - PLlacO – promoter,  BBa_R0011: the promoter show the highest RPU-value of all characterized promoters and had the highest with IPTG.
+. Main Page (http://partsregistry.org/Part:BBa_J23113) - constitutive promoter family member, BBa_J23113
+Parts J23100 through J23119 are a family of constitutive promoter parts isolated from a small combinatorial library. As expected J23113 showed a low promoter strength.
+. Main page http://partsregistry.org/Part:BBa_K592026 J23101-B0032-BFP, BBa_K592026: the constitutive promoter was used as the reference in the characterization measurement.
+<div class="grid_Orange_thin omega mywhite">
+Characterized Parts
+</div>
+Our measurement with the FACS AriaII shown the following  Relative Promoter Units with the Blue Fluorescent Protein (mTagBFP) as reporter, assembled with B0032 in pSB3K3-backbone transformed in TOP10 E.coli-strain.
+[[image:Promoter_characterization.PNG|400px]]
+. Main page http://partsregistry.org/Part:BBa_K592003  - PcpcG2 promoter, BBa_K592003: measurement shows a RPU-value of 0,1.
+. Main page http://partsregistry.org/Part:BBa_K592006 - PFixK2 promoter, BBa_K592006 is activated by the blue sensor and showed no significant expression in absence of the YF1-sensor (BBa_K592004)
+. Main page http://partsregistry.org/Part:BBa_K592008 -  PT5-lac Promoter, BBa_K592008: showed high expression of BFP, and showed higher expression in the presence of  IPTG than without it.
 {{Template:Uppsala-SwedenTemplatefooter}}