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Team:Evry/auxin production
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Revision as of 16:44, 24 September 2012
Auxin production
Overview
The 2011 Imperial College iGEM [1] project showed that the plant hormone auxin or indole-3-acetic acid (IAA) can be synthetized in Escherichia coli through a two-step pathway from tryptophan. In their work, the genes encoding the auxin biosynthesis pathway originally coming from Pseudomonas savastanoi were expressed in E. coli. In our project, we aim to express this auxin biosynthesis pathway in Xenopus tropicalis . First, we plan to reengineer the auxin biosynthesis pathway in the tadpole with a constitutive and ubiquitous promoter. Once the system is functional, we intend to put the system under the control of inducible and tissue-specific promoters.
By modelling the auxin biosynthesis pathway, we will be able to determine the number of produced auxin molecules per plasmids injected in the cell, as well as, the number of diffused auxin molecules through the plasma membrane into the extracellular medium.
Assumptions
-
Model description
The plasmids containing the two genes involved in the auxin biosynthesis pathway, namely iaaM and iaaH, will be expressed in a constitutive and ubiquitous manner in a first step and in a second step in an inducible and tissue-specific manner. The iaaM gene encodes tryptophan-2-monooxygenase (IAAM) that catalyses the conversion of tryptophan (Trp) into indole-3-acetamide (IAM). The iaaH gene encodes indoleacetamide hydrolase (IAAH) that hydrolyse IAM to realease indole-3-acetic acid (IAA) [2]. At the same time, the synthesized IAM and IAA will competitively inhibit the enzyme activity of IAAM.
All the reactions involved in the auxin biosynthesis pathway are illustrated in Figure 1.
Figure 1. Auxin synthesis pathway (Diagram by 2012 Évry iGEM team)
Equations
![]()
where:
-
Limit conditions
Sensitivity analysis
Results
Conclusion
References:
- https://2011.igem.org/Team:Imperial_College_London
- Cheng, Y. Dai, C. Zhao, Y. 2006. Auxin biosynthesis by the YUCCA flavin monooxygenases controls the formation of floral organs and vascular tissues in Arabidopsis. Genes & Dev 20: 1790-1799. Doi: 10.1101/gad.1415106
Assumptions
Model description
The plasmids containing the two genes involved in the auxin biosynthesis pathway, namely iaaM and iaaH, will be expressed in a constitutive and ubiquitous manner in a first step and in a second step in an inducible and tissue-specific manner. The iaaM gene encodes tryptophan-2-monooxygenase (IAAM) that catalyses the conversion of tryptophan (Trp) into indole-3-acetamide (IAM). The iaaH gene encodes indoleacetamide hydrolase (IAAH) that hydrolyse IAM to realease indole-3-acetic acid (IAA) [2]. At the same time, the synthesized IAM and IAA will competitively inhibit the enzyme activity of IAAM. All the reactions involved in the auxin biosynthesis pathway are illustrated in Figure 1.
Figure 1. Auxin synthesis pathway (Diagram by 2012 Évry iGEM team)
Equations
Limit conditions
Sensitivity analysis
Results
Conclusion
References:
- https://2011.igem.org/Team:Imperial_College_London
- Cheng, Y. Dai, C. Zhao, Y. 2006. Auxin biosynthesis by the YUCCA flavin monooxygenases controls the formation of floral organs and vascular tissues in Arabidopsis. Genes & Dev 20: 1790-1799. Doi: 10.1101/gad.1415106
