Team:LMU-Munich/Results

From 2012.igem.org

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===''B. subtilis'' vectors===
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===Seven novel vectors were constructed and four already proven to work in ''B. subtilis''.===
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===15 Promoters evaluated in ''B. subtilis'' and added to the registry!===
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<br>
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====Overview of all evaluated promoters====
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<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>
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<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>
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<br>
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{| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
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<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>
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<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>
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===Sporobeads - what protein do you want to display?===
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|[[File:Fluorescence of Sporobeads.png|610px|center]]
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<font color="#000000"; size="2">Result of fluorescence evaluation of the three strains: W168, B53 and B70.</font>
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===Germination Gene Knockouts===
 
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===Erase<sup>*</sup> germination ability! Knockout strains===
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{| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
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|[[File:Germination_RatesII.jpg|600px|center]]
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|[[File:Germination_RatesIIii_ii.jpg|600px|center]]
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<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 />
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<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 />
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'''WT168''': Wild-type 168 <br />
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'''W168''': wild-type 168 <br />
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'''''spo0A''::tet''': A sporulation-deficient strain <br />
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'''''spo0A''::tet''': a sporulation-deficient strain <br />
'''B29''': ''cwlD''::kan, ''sleB''::mls <br />
'''B29''': ''cwlD''::kan, ''sleB''::mls <br />
'''B30''': ''gerD''::cm, ''sleB''::mls <br />
'''B30''': ''gerD''::cm, ''sleB''::mls <br />
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'''B43''': ''gerD''::cm, ''sleB''::mls, ''cwlJ''::spec <br />
'''B43''': ''gerD''::cm, ''sleB''::mls, ''cwlJ''::spec <br />
'''B46''': ''cwlD''::kan, ''cwlJ''::spec, ''gerD''::cm, ''sleB''::mls <br />
'''B46''': ''cwlD''::kan, ''cwlJ''::spec, ''gerD''::cm, ''sleB''::mls <br />
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'''B47''': ''gerD''::cm, ''sleB''::mls, ''cwlJ''::spec, ''cwlD''::kan</font>
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'''B47''': ''gerD''::cm, ''sleB''::mls, ''cwlJ''::spec, ''cwlD''::kan<br></font>
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<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>
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====Inverter====
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===Inverter works!===
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[[File:LMU Inverter graph.png|600px]]
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|[[File:LMU Inverter graph.png|600px|center]]
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<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>
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For detailed results visit our [[Team:LMU-Munich/Data|data page]] and for all background info our [[Team:LMU-Munich/Project|project page]].
For detailed results visit our [[Team:LMU-Munich/Data|data page]] and for all background info our [[Team:LMU-Munich/Project|project page]].
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Latest revision as of 22:13, 26 September 2012

iGEM Ludwig-Maximilians-Universität München Beadzillus

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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".

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