Team:Tianjin/Project/Experiment/TetA

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                        <a href="https://2013.igem.org/Team:Tianjin/Project/Experiment/Endo-Induce">Endogenous Alkanes Induce</a>
 
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                        <a href="https://2013.igem.org/Team:Tianjin/Project/Experiment/TetA">TetA</a>
 
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<center><span style="font-family:Arial;font-size:46px;color:#000;"> Experiment</span></center>
 
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=Endogenous Induce&Resistance Selection=
 
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<p>Here, we want to test the resistance against tetracycline of Alk-Sensor induced by endogenous alkanes.</p>
 
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<p>We put the alkane producing module into cells to test the function of TetA:</p>
 
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<div style="text-align:center;vertical-align:middle;"><a href="#" target="_blank" ><img src="#" width="500px" /></a></div>
 
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<p><b>Construct:</b></p>
 
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<p>Left: No.60: TetA downstream of Palkm and AlkR downstream of J23103, are cloned into vector PSB1C3 and there’s a alkane producing vector PSB3K3.</p>
 
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<p>Right: No.61: We construct a blank vector PSB3K3 without NPDC and AAR as blank control, and the other vector PSB1C3 is the same as No.60.</p>
 
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<p><b>Characterization: </b></p>
 
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<p>Strains of No.60 are cultured in LB medium for about 12 hours. Add 17μg/mL Kan, 12μg/mL Cm and 12μg/mL Tc respectively. After that, inoculate into 3 ml LB medium for an overnight cultures at 37 ℃ with 220rpm shaking.</p>
 
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<p><b>Result:</b> </p>
 
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<p>Alkane producing E.coli can grow in tube but blank vector E.coli cannot grow .</p>
 
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<p><b>Conclusion:</b></p>
 
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<p>Synthesized alkanes induce the expression of alkR, which enables E.coli to survive under the pressure of Tetracycline.</p>
 
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<p>Thus, we can get a 3×3 matrix, with promoter strength from weak, medium to strong, and plasmid copy number from small, medium to large.</p>
 
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Latest revision as of 02:27, 29 October 2013

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