Team:LMU-Munich/Results

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Latest revision as of 22:13, 26 September 2012

The LMU-Munich team is exuberantly happy about the great success at the World Championship Jamboree in Boston. Our project Beadzillus finished 4th and won the prize for the "Best Wiki" (with Slovenia) and "Best New Application Project".

[ more news ]

Main Results banner.jpg

Here you will find the main results of our work.

Seven novel vectors were constructed and four already proven to work in B. subtilis.

Name BioBrick	E. coli res.	B. subt. res.	Insertion locus	Description	Cloning status	Works?
pSB_Bs1C [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823023 (BBa_K823023)]	Amp	Cm	amyE	empty
pSB_Bs4S [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823022 (BBa_K823022)]	Amp	Spec	thrC	empty
pSB_Bs2E [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823027 (BBa_K823027)]	Amp	MLS	lacA	empty	in work	not tested
pSB_Bs1C-lacZ [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823021 (BBa_K823021) ]	Amp	Cm	amyE	lacZ reporter
pSB_Bs3C-luxABCDE [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823025 (BBa_K823025)]	Amp	Cm	sacA	luxABCDE reporter
pSB_Bs4S-P_Xyl [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823024 (BBa_K823024)]	Amp	Spec	thrC	Xylose-promoter		refuses transformation
pSB_Bs0K-P_spac [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823026 (BBa_K823026)]	Amp	Kan	replicative	IPTG-promoter		unexpected
Sporovector [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823026 (BBa_K823054)]	Amp	Spec	thrC	to make Sporobeads		not tested

15 Promoters evaluated in B. subtilis and added to the registry!

This section gives an overview on the strength of all evaluated promoters, which span a large range of activities. For more details and informations of the experiments see the Data page of the promoters. Note that P_veg was not evaluated with luminescence measurements and this bar is just projected from the results of the β-galactosidase assay.

Overview of promoter activity evaluated with luminescence measurements in pSB_Bs3C-luxABCDE. These values derive from the experiments you can find in our Data section. Lumi per OD₆₀₀ are taken at a OD₆₀₀ of 0.1. Values are the average and the standard deviation of three different experiments for clone 1. Shown is the activity of the Anderson promoters J23100 (#100), J23101 (#101), J23102 (#102), J23103 (#103), J23106 (#106), J23107 (#107), J23113 (#113), J23114 (#114), J23115 (#115), J23117 (#117), J23118 (#118) as well as the activity of the constitutive promoters P_liaG, and P_lepA. The activity of the inducible promoter P_liaI is shown with (+bac) and without (-bac) induction with bacitracin (10 μg/ml). The promoter activity of P_veg is projected from the results from the β-galactosidase assay and was not measured with luminescence measurements. For the assays shown, an intermediate version of the reporter vector pSB_Bs3C-luxABCDE was used, which still contained one forbidden PstI site.

Sporobeads - what protein do you want to display?

Result of fluorescence evaluation of the three strains: W168, B53 and B70.

Erase^* germination ability! Knockout strains

The germination rates of B. subtilis germination mutants. All germination rates are relative to wild type 168 (W168), our positive control. W168 showed near 100% germination rates. The mutant spo0A::tet is unable to form spores, and should therefore show no germination (our negative control). Our results were consistent with this expectation. Mutants used were:
W168: wild-type 168
spo0A::tet: a sporulation-deficient strain
B29: cwlD::kan, sleB::mls
B30: gerD::cm, sleB::mls
B32: cwlJ::spec, cwlD::kan
B40: cwlD::kan, sleB::mls, cwlJ::spec
B41: cwlD::kan, sleB::mls, gerD::cm
B42: cwlD::kan, cwlJ::spec, gerD::cm
B43: gerD::cm, sleB::mls, cwlJ::spec
B46: cwlD::kan, cwlJ::spec, gerD::cm, sleB::mls
B47: gerD::cm, sleB::mls, cwlJ::spec, cwlD::kan

*No germinating spores detected in 4.6x10⁹ spores. For a full explanation of these great results, see All results.

Inverter works!

β-Galactosidase assay of Inverter with lacZα as reporter. The higher the arabinose concentration, the higher the translational repression of uof_CGU-lacZα

For detailed results visit our data page and for all background info our project page.

Project Navigation


Bacillus Intro	Bacillus BioBrickBox	Sporobeads	Germination STOP

Retrieved from "http://2012.igem.org/Team:LMU-Munich/Results"

@@ Line 4: / Line 4: @@
 Here you will find the main results of our work.
-===''B. subtilis'' vectors===
+===Seven novel vectors were constructed and four already proven to work in ''B. subtilis''.===
+<br>
 {| class="colored"
 !Name
@@ Line 82: / Line 84: @@
-====Overview of all evaluated promoters====
+===15 Promoters evaluated in ''B. subtilis'' and added to the registry!===
+<br>
-<p align="justify"> This section gives an overview of all evaluated promoters which cover a large range of activity. For more details and informations of the experiments see the [https://2012.igem.org/Team:LMU-Munich/Data Data] page of the promoters. Note that P<sub>''veg''</sub> was not evaluated with luminescence measurements and this bar is just projected from the results of the beta-galactosidase assay.</p>
+<p align="justify"> This section gives an overview on the strength of all evaluated promoters, which span a large range of activities. For more details and informations of the experiments see the [https://2012.igem.org/Team:LMU-Munich/Data Data] page of the promoters. Note that P<sub>''veg''</sub> was not evaluated with luminescence measurements and this bar is just projected from the results of the β-galactosidase assay.</p>
 <br>
 {| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
 | style="width: 70%;background-color: #EBFCE4;" |
@@ Line 95: / Line 96: @@
 {| style="color:black;" cellpadding="0" width="100%" cellspacing="0" border="0" align="center" style="text-align:center;"
 |style="width: 70%;background-color: #EBFCE4;" |
-<font color="#000000"; size="2"><p align="justify"> '''Overview of promoter activity evaluated with luminescence measurements in pSB<sub>''Bs''</sub>3C-''luxABCDE''.''' These values derive from the experiments you can find in our Data section. Lumi per OD<sub>600</sub> are taken at a OD<sub>600</sub> of 0.1. Values are the average and the standard deviation of three different experiments for clone 1. Shown is the activity of the Anderson promoters J23100 (#100), J23101 (#101), J23102 (#102), J23103 (#103), J23106 (#106), J23107 (#107), J23113 (#113), J23114 (#114), J23115 (#115), J23117 (#117), J23118 (#118) as well as the activity of the constitutive promoters P<sub>''liaG''</sub>, and P<sub>''lepA''</sub>. The activity of the inducible promoter P<sub>''liaI''</sub> is shown with (+bac) and without (-bac) induction with bacitracin (10 μg/ml). The promoter activity of P<sub>''veg''</sub> is projected from the results from the beta-galactosidase assay and was not measured with luminescence measurements. </p></font>
+<font color="#000000"; size="2"><p align="justify"> '''Overview of promoter activity evaluated with luminescence measurements in pSB<sub>''Bs''</sub>3C-''luxABCDE''.''' These values derive from the experiments you can find in our Data section. Lumi per OD<sub>600</sub> are taken at a OD<sub>600</sub> of 0.1. Values are the average and the standard deviation of three different experiments for clone 1. Shown is the activity of the Anderson promoters J23100 (#100), J23101 (#101), J23102 (#102), J23103 (#103), J23106 (#106), J23107 (#107), J23113 (#113), J23114 (#114), J23115 (#115), J23117 (#117), J23118 (#118) as well as the activity of the constitutive promoters P<sub>''liaG''</sub>, and P<sub>''lepA''</sub>. The activity of the inducible promoter P<sub>''liaI''</sub> is shown with (+bac) and without (-bac) induction with bacitracin (10 μg/ml). The promoter activity of P<sub>''veg''</sub> is projected from the results from the β-galactosidase assay and was not measured with luminescence measurements. For the assays shown, an intermediate version of the reporter vector pSB<sub>''Bs''</sub>3C-''luxABCDE'' was used, which still contained one forbidden PstI site.</p></font>
 |}
 |}
@@ Line 101: / Line 102: @@
+===Sporobeads - what protein do you want to display?===
+<br>
+{| style="color:black;" cellpadding="3" width="100%" cellspacing="0" border="0" align="center" style="text-align:left;"
+| style="width: 70%;background-color: #EBFCE4;" |
+{|
+|[[File:Fluorescence of Sporobeads.png|610px|center]]
+|}
+|-
+| style="width: 70%;background-color: #EBFCE4;" |
+{| style="color:black;" cellpadding="0" width="100%" cellspacing="0" border="0" align="center" style="text-align:center;"
+|style="width: 70%;background-color: #EBFCE4;" |
+<font color="#000000"; size="2">Result of fluorescence evaluation of the three strains: W168, B53 and B70.</font>
+|}
+|}
-===Germination Gene Knockouts===
+===Erase<sup>*</sup> germination ability! Knockout strains===
+<br>
 {| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
 | style="width: 70%;background-color: #EBFCE4;" |
 {|align:center
-|[[File:Germination_RatesII.jpg|600px|center]]
+|[[File:Germination_RatesIIii_ii.jpg|600px|center]]
 |-
 | style="width: 80%;background-color: #EBFCE4;" |
 {| style="color:black;" cellpadding="3" width="95%" cellspacing="0" border="0" align="left" style="text-align:left;"
 |style="width: 70%;background-color: #EBFCE4;" |
-<font color="#000000"; size="2">'''The germination rates of ''B. subtilis'' germination mutants.''' All germination rates are relative to wild type 168 (WT168), our positive control. The mutant ''spo0A''::tet is unable to form spores, and should therefore show no germination (our negative control). Mutants used were: <br />
+<font color="#000000"; size="2">'''The germination rates of ''B. subtilis'' germination mutants.''' All germination rates are relative to wild type 168 (W168), our positive control. W168 showed near 100% germination rates. The mutant ''spo0A''::tet is unable to form spores, and should therefore show no germination (our negative control). Our results were consistent with this expectation. Mutants used were: <br />
-'''WT168''': Wild-type 168 <br />
+'''W168''': wild-type 168 <br />
-'''''spo0A''::tet''': A sporulation-deficient strain <br />
+'''''spo0A''::tet''': a sporulation-deficient strain <br />
 '''B29''': ''cwlD''::kan, ''sleB''::mls <br />
 '''B30''': ''gerD''::cm, ''sleB''::mls <br />
@@ Line 123: / Line 139: @@
 '''B43''': ''gerD''::cm, ''sleB''::mls, ''cwlJ''::spec <br />
 '''B46''': ''cwlD''::kan, ''cwlJ''::spec, ''gerD''::cm, ''sleB''::mls <br />
-'''B47''': ''gerD''::cm, ''sleB''::mls, ''cwlJ''::spec, ''cwlD''::kan</font>
+'''B47''': ''gerD''::cm, ''sleB''::mls, ''cwlJ''::spec, ''cwlD''::kan<br></font>
 |}
 |}
 |}
+<p align="justify">*No germinating spores detected in 4.6x10<sup>9</sup> spores. For a full explanation of these great results, see [https://2012.igem.org/Team:LMU-Munich/Data/Knockout All results].</p>
+===Inverter works!===
+<br>
+{| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
+| style="width: 70%;background-color: #EBFCE4;" |
+{|
+|[[File:LMU Inverter graph.png|600px|center]]
+|-
+| style="width: 70%;background-color: #EBFCE4;" |
+{| style="color:black;" cellpadding="0" width="100%" cellspacing="0" border="0" align="center" style="text-align:center;"
+|style="width: 70%;background-color: #EBFCE4;" |
+<font color="#000000"; size="2"><p align="justify"> β-Galactosidase assay of Inverter with ''lacZα'' as reporter. The higher the arabinose concentration, the higher the translational repression of ''uof<sub>CGU</sub>-lacZα'' </p></font>
+|}
+|}
+|}
-====Inverter====
-[[File:LMU Inverter graph.png|600px]]
 For detailed results visit our [[Team:LMU-Munich/Data|data page]] and for all background info our [[Team:LMU-Munich/Project|project page]].
-</div>
 <div class="box">
@@ Line 150: / Line 181: @@
 |[[Team:LMU-Munich/Germination_Stop|<font size="2" face="verdana">'''Germination'''<BR>STOP</font>]]
 |}
+</div>