Difference between revisions of "Part:BBa K584000:Experience"
 
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===Applications of BBa_K584000===
 
===Applications of BBa_K584000===
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'''Group:'''iGEM_BIT2017
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'''Author:'''WENJIA WANG
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We transformed the part BBa_K584000 into DH5α E.coli and tested its OD600 and GFP fluorescence. L-arabinose was added with the amount of 0mg，40.5mg，91.0mg, and 121.5mg respectively. The results show that the OD600 and GFP fluorescence grows obviously with the concentration gradient of the L-arabinose. (Fig.1;Fig.2)
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[[Image: BIT Figure copper 3 wwj.png |thumb|center|500px|Fig.1]]
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[[Image: BIT Figure copper 4 wwj.png |thumb|center|500px|Fig.2]]
  
 
===User Reviews===
 
===User Reviews===
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===User Reviews===
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<partinfo>BBa_K584000 AddReview number</partinfo>
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<partinfo>BBa_K584001 AddReview 5</partinfo>
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<I>Username</I>
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<I><B>[http://2011.igem.org/Team:KULeuven K.U.Leuven iGEM 2011 Team]</B></I>
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Enter the review inofrmation here.
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<u><b>Characterization</b></u>
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<p align="justify">To test the usefulness of the arabinose-inducible promoter I13453 in our 2011 iGEM project, we fused the promoter to a GFP reporter, and assayed the promoter’s activity after addition of different amounts of arabinose. We tested this activity both in a TOP10F’ (Figure 1) as well as a MG1655 (Figure 2) E.coli strain background. For more information on E.coli strain descriptions, we recommend the following web site: [http://openwetware.org/wiki/E._coli_genotypes E.Coli_Genotypes].</p>
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[[Image:pBAD_TOP10.jpg|thumb|center|Figure1]]
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<p align="justify">As can be seen in Figure 1, addition of arabinose had only minor, if any, effect on the growth of TOP10F’ cells, most likely because these cells cannot metabolize the sugar. However, to our surprise, we did not observe an arabinose-induced increase of fluorescence compared to the control without arabinose for the first 6 hours. The observed increase in fluorescence after arabinose addition at the 8 hour time point could not be confirmed when the experiment was repeated.</p>
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[[Image:pBAD_MG1655.jpg|thumb|center|Figure2]]
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<p align="justify">Figure 2 demonstrates that in MG1655 cells, the addition of 0.2% and 2% arabinose results in a growth defect after about 4 hours of growth, which may relate to the metabolism of the sugar in this strain. In contrast to the TOP10F’ cells, we see a clear induction of promoter activity after addition of arabinose. The addition of only 0.02% resulted in the greatest induction. Increasing arabinose concentration did not increase fluorescence, probably due to the observed growth defect. </p>
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<p align="justify">As an additional control, we checked the activity of a constitutive promoter under the same conditions as described here. For results on these experiments, check out our [https://parts.igem.org/wiki/index.php?title=Part:BBa_K584001 BBa_K584001] page.</p>
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Latest revision as of 00:17, 28 October 2017

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Please enter how you used this part and how it worked out.

Applications of BBa_K584000

Group:iGEM_BIT2017

Author:WENJIA WANG

We transformed the part BBa_K584000 into DH5α E.coli and tested its OD600 and GFP fluorescence. L-arabinose was added with the amount of 0mg，40.5mg，91.0mg, and 121.5mg respectively. The results show that the OD600 and GFP fluorescence grows obviously with the concentration gradient of the L-arabinose. (Fig.1;Fig.2)

Fig.1
Fig.2

User Reviews

User Reviews

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