Difference between revisions of "Part:BBa K2753002"
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<partinfo>BBa_K2753002 short</partinfo>
 
<partinfo>BBa_K2753002 short</partinfo>
  
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<p>GPPS: Geranyl diphosphate (GPP), the entry point to the formation of terpene moiety, is a product of the condensation of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) by GPP synthase (GPPS)<sup>[1]</sup>. IPP and DMAPP are supplied by either MVA pathway or MEP pathway or both depending on species. This parts originated from the species Abies Grandis, and it’s codon-optimized for the use in E. coli.</p></br>
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<p>GPPS: Geranyl diphosphate (GPP), the entry point to the formation of terpene moiety, is a product of the condensation of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) by GPP synthase (GPPS)<sup>[1]</sup>. IPP and DMAPP are supplied by either MVA pathway or MEP pathway or both depending on species. This parts originated from the species Abies Grandis, and it’s codon-optimized for the use in E. coli.</p>
  
 
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<center>
 
<Figure>
 
<Figure>
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<img width="70%" src="http://2018.igem.org/File:T--GreatBay_China--_GPPS_pathway.png">
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<img width="70%" src="https://static.igem.org/mediawiki/2018/9/97/T--GreatBay_China--_GPPS_pathway.png">
 
<figcaption>Fig.1 The biosynthesis pathway for GPP <sup>[2]</sup>.</figcaption>
 
<figcaption>Fig.1 The biosynthesis pathway for GPP <sup>[2]</sup>.</figcaption>
 
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===Characterization===
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<p>In our study, to investigate the optimal expression level of GPPS and GES for geraniol production, we assembled GPPS and obGES to make a geraniol-generating operon without a promoter(Part#), We employed three different promoters: </p><br>
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<ul>
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<li>pTALE sp1 (https://parts.igem.org/Part:BBa_K2753000), copy number-independent promoter of medium strength</li>
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<li>pTALE sp2 (https://parts.igem.org/Part:BBa_K2753001), copy number-independent promoter of low strength</li>
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<li>pTac (https://parts.igem.org/Part:BBa_K180000), IPTG inducible promoter</li>
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</ul>
  
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<p>And three vectors of unique copy number:</p><br>
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<ul>
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<li>pUC20: ~500</li>
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<li>pR6K: ~15</li>
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<li>pSC101: ~1<li>
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</ul>
  
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<!-- Add more about the biology of this part here
 
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===Usage and Biology===
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2753002 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2753002 SequenceAndFeatures</partinfo>

Revision as of 10:00, 1 October 2018


pSB1C3-GPPS cds

GPPS: Geranyl diphosphate (GPP), the entry point to the formation of terpene moiety, is a product of the condensation of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) by GPP synthase (GPPS)[1]. IPP and DMAPP are supplied by either MVA pathway or MEP pathway or both depending on species. This parts originated from the species Abies Grandis, and it’s codon-optimized for the use in E. coli.

Fig.1 The biosynthesis pathway for GPP [2].

Characterization

In our study, to investigate the optimal expression level of GPPS and GES for geraniol production, we assembled GPPS and obGES to make a geraniol-generating operon without a promoter(Part#), We employed three different promoters:


And three vectors of unique copy number:


  • pUC20: ~500
  • pR6K: ~15
  • pSC101: ~1

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Referece

[1]: Rai, A., Smita, S. S., Singh, A. K., Shanker, K., & Nagegowda, D. A. (2013). Heteromeric and Homomeric Geranyl Diphosphate Synthases from Catharanthus roseus and Their Role in Monoterpene Indole Alkaloid Biosynthesis. Molecular Plant, 6(5), 1531–1549. doi:10.1093/mp/sst058

[2]: Zebec, Z., Wilkes, J., Jervis, A. J., Scrutton, N. S., Takano, E., & Breitling, R. (n.d.). Towards synthesis of monoterpenes and derivatives using synthetic biology. Current Opinion in Chemical Biology, 34, 37–43. https://doi.org/10.1016/j.cbpa.2016.06.002