Difference between revisions of "Part:BBa K2062005:Experience"

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  <h2>Transformation of <em>P. putida</em> KT2440</h2>
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  <p>
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This experience page is provided so that any user may enter their experience using this part.<br>
    In order to avoid the virulence factors of
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Please enter how you used this part and how it worked out.
    <em>Pseudomonas aeruginosa</em>, bacterial strains with
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    similar or shared metabolic pathways to the one
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    above were chosen as potential candidates. The
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    final candidates were <em>Pseudomonas putida</em> and
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    <em>Staphylococcus epidermidis</em>. Although
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    <em>S. epidermidis</em> doesn’t share the same exact
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    pathway as <em>P. aeruginosa</em>, it is a
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    naturally-occurring skin microbiome and only need
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    two additional enzymes, RhlA and RhlB, to produce
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    mono-rhamnolipids. Genes rhlA and rhlB necessary
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    for mono-rhamnolipid synthesis were extracted from
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    the <em>P. aeruginosa P14</em> bacterial strain. These
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    genes were cloned into the modified plasmid pNJ3.1
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    using standard cloning methods for transformation
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    into the desired bacterial strains (Figure 2). The
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    plasmid pC194 and a shuttle vector strain,
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    <em>S. aureus</em> RN4220 (details on <em>S. epidermidis</em>
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    transformation are discussed in the experiments
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    and result section) were used for <em>S. epidermidis</em>
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    transformations with the same basic design (Figure
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    3). The conversion of mono-rhamnolipids to
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    di-rhamnolipids requires the additional gene rhlC,
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    which was also extracted from P14 strain and
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    cloned into the same pNJ3.1 vector (Figure 4).
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  </p>
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  <figure>
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    <img src="https://static.igem.org/mediawiki/parts/a/a4/RhlAB_circuit.png"
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alt="RhlAB Plasmid Circuit" width="500">
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  </figure>
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===Applications of BBa_K2062005===
 
===Applications of BBa_K2062005===
  

Latest revision as of 04:14, 24 October 2016


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Applications of BBa_K2062005

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