Difference between revisions of "Part:BBa K1602017"
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         <img class="shrinkToFit transparent" alt="https://static.igem.org/mediawiki/parts/f/f1/TU_Darmstadt_EG_XylB-xylC.png" src="https://static.igem.org/mediawiki/parts/f/f1/TU_Darmstadt_EG_XylB-xylC.png" height="407" width="169">
 
         <img class="shrinkToFit transparent" alt="https://static.igem.org/mediawiki/parts/f/f1/TU_Darmstadt_EG_XylB-xylC.png" src="https://static.igem.org/mediawiki/parts/f/f1/TU_Darmstadt_EG_XylB-xylC.png" height="407" width="169">
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             <li><b>Figure 1</b> Sheme of the reactions catalyzed by xylB and xylC. The xylC reaction can also happen spontaneously but in a much lower speed.</li>
 
             <li><b>Figure 1</b> Sheme of the reactions catalyzed by xylB and xylC. The xylC reaction can also happen spontaneously but in a much lower speed.</li>
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===<h2>Results</h2>===
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The expression of cadA has been visualized via SDS-PAGE. Positive clones were grown at 37&deg; celsius until an OD of 0,7. Afterwards the cells were induced utilizing 20&micro;l of 1M IPTG for 12h at 28&deg; celsius. Finally the cells were lysated via ultrasonic cell disruption.
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                <div><img style="width: 200px; height: 389px;" src="https://static.igem.org/mediawiki/2015/3/35/Da15_sds_cadA.png" href="https://static.igem.org/mediawiki/2015/3/35/Da15_sds_cadA.png"></div>
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                <div><img style="width: 300px; height: 456;" src="https://static.igem.org/mediawiki/2015/a/a7/Da15_sds_cadA_plot.png" href="https://static.igem.org/mediawiki/2015/a/a7/Da15_sds_cadA_plot.png"</div>
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                <div><b>Figure 3</b> Scan of the PAGE containing from left to right a marker (M; Protein Marker III AppliChem), the positive sample (1) and a negative control (2). The picture was cropped and edited for clarification purposes.</div>
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                <div><b>Figure 4</b> Plot of the gel lanes based on contrast analyses - created with ImageJ</div>
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===<h2>Sequence and Features</h2>===
 
===<h2>Sequence and Features</h2>===

Revision as of 20:32, 18 September 2015

D-xylonic acid producing operon

D-Xylose is a monosaccharide belonging to the aldopentose family. It was recently shown that the D-xylose dehydrogenase xylB from Caulobacter crescentus can convert D-xylose to D-xylonolactone. This can react spontaneously or through the catalysation of xylC to D-xylonic acid. In E. coli D-xylonic acid can be further metabolized to ethyleneglycol.


https://static.igem.org/mediawiki/parts/f/f1/TU_Darmstadt_EG_XylB-xylC.png

  • Figure 1 Sheme of the reactions catalyzed by xylB and xylC. The xylC reaction can also happen spontaneously but in a much lower speed.



    • Usage

    This part is a composite of two coding genes with strong RBS (BBa_B0034). The transcription is controlled by a T7 promotor (BBa_I719005).


    Figure 2



    Results

    The expression of cadA has been visualized via SDS-PAGE. Positive clones were grown at 37° celsius until an OD of 0,7. Afterwards the cells were induced utilizing 20µl of 1M IPTG for 12h at 28° celsius. Finally the cells were lysated via ultrasonic cell disruption.

    Figure 3 Scan of the PAGE containing from left to right a marker (M; Protein Marker III AppliChem), the positive sample (1) and a negative control (2). The picture was cropped and edited for clarification purposes.
    Figure 4 Plot of the gel lanes based on contrast analyses - created with ImageJ

    ===

    Sequence and Features

    === BBa_K1602017 SequenceAndFeatures
    ===References=== 1. Liu H, Ramos KR, Valdehuesa KN, Nisola GM, Lee WK, Chung WJ. Biosynthesis of ethylene glycol in Escherichia coli. Appl Microbiol Biotechnol. 2013;97(8):3409-17. 2. Toivari MH, Nygard Y, Penttila M, Ruohonen L, Wiebe MG. Microbial D-xylonate production. Appl Microbiol Biotechnol. 2012;96(1):1-8.