Difference between revisions of "Part:BBa K515100"
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<partinfo>BBa_K515100 short</partinfo>
 
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<h2>Background</h2>
 
<p> The IAM pathway is a two step pathway which generates indole-3-acetic acid (IAA), also known as auxin, from the precursor tryptophan. IAA tryptophan monooxygenase (IaaM) <a href="https://parts.igem.org/Part:BBa_K515000">BBa_K515000</a>,  catalyzes the oxidative carboxylation of L-tryptophan to indole-3-acetamide which is hydrolyzed to indole-3-acetic acid and ammonia by indoleacetamide hydrolase (IaaH) <a href="https://parts.igem.org/Part:BBa_K515001">BBa_K515001</a> . There are several different pathways that produce indole-3-acetic acid.[1]
 
<p> The IAM pathway is a two step pathway which generates indole-3-acetic acid (IAA), also known as auxin, from the precursor tryptophan. IAA tryptophan monooxygenase (IaaM) <a href="https://parts.igem.org/Part:BBa_K515000">BBa_K515000</a>,  catalyzes the oxidative carboxylation of L-tryptophan to indole-3-acetamide which is hydrolyzed to indole-3-acetic acid and ammonia by indoleacetamide hydrolase (IaaH) <a href="https://parts.igem.org/Part:BBa_K515001">BBa_K515001</a> . There are several different pathways that produce indole-3-acetic acid.[1]
 
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IaaM and IaaH originate from <i>P.savastanoi</i> and have been expressed in <i>E. coli</i> previously, and shown to secrete auxin into cell supernatant.[2]</p>  
 
IaaM and IaaH originate from <i>P.savastanoi</i> and have been expressed in <i>E. coli</i> previously, and shown to secrete auxin into cell supernatant.[2]</p>  
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<h2>Experimental Data</h2>
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<img src="https://static.igem.org/mediawiki/2011/a/ad/ICL_Latest.standard.curve.png" width=500px/>
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<p><i>Figure 1: Standard curve of Salkowski assay made with synthetic IAA in LB</i></p>
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<img src="https://static.igem.org/mediawiki/parts/5/58/ICL_salkowski_cuvettes.JPG" width=300px/>
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<p><i>Figure 2: Cuvettes used to measure OD for the standard curve. As IAA concentration increases, the solution progresses towards red. </i></p>
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<!-- Add more about the biology of this part here
 
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Revision as of 12:46, 21 September 2011

IAA biosynthetic genes under control of the Pveg2 promoter

Background

The IAM pathway is a two step pathway which generates indole-3-acetic acid (IAA), also known as auxin, from the precursor tryptophan. IAA tryptophan monooxygenase (IaaM) BBa_K515000, catalyzes the oxidative carboxylation of L-tryptophan to indole-3-acetamide which is hydrolyzed to indole-3-acetic acid and ammonia by indoleacetamide hydrolase (IaaH) BBa_K515001 . There are several different pathways that produce indole-3-acetic acid.[1]

IaaM and IaaH originate from P.savastanoi and have been expressed in E. coli previously, and shown to secrete auxin into cell supernatant.[2]

Experimental Data

Figure 1: Standard curve of Salkowski assay made with synthetic IAA in LB

Figure 2: Cuvettes used to measure OD for the standard curve. As IAA concentration increases, the solution progresses towards red.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 547
    Illegal BamHI site found at 1492
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 254
    Illegal NgoMIV site found at 2835
  • 1000
    COMPATIBLE WITH RFC[1000]


References

[1]Spaepen S. et al., 2007. Indole-3-acetic acid in microbial and microorganism-plant signaling. Federation of European Microbiological Societies Microbiology Reviews , 31, pp.425–448.

[2]Palm, CJ et al., 1989. Cotranscription of genes encoding indoleacetic acid production in Pseudomonas syringae subsp. savastanoi. Journal of Bacteriology, 171(2), pp.1002-1009.