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Revision as of 12:34, 17 September 2015

IGEM Bordeaux 2015



Parts

Our submitted genes are listed down below

<groupparts>iGEM015 Bordeaux</groupparts>


Curdlan synthase gene CrdS with codon optimization for E. coli : BBa_K1686046

Curdlan is a bacterial polysaccharide (linear (1->3)-β-glucan) that has been of significant recent interest due to its valuable properties and its bioactivity. In fact, Curdlan belongs to the class of biological response modifiers that enhance or restore normal immune defenses. For example, it can have antitumor, anti-infective, anti-inflammatory, and anticoagulant activities (see other properties of Curdlan). In particular, this (1->3)-β-glucan can stimulate plants' immune system. Our project is focused on the production of curdlan for stimulate plant defense system against Downy Mildew.
The molecular genetics of curdlan production have been investigated in Agrobacterium sp. ATCC31749. Studies identified three genes (crdA, crdS and crdC) that are essential for curdlan production The crdASC genes occupy a contiguous 4,948 bp region of the genome ( the crdS gene is flanked by the crdA and crdC genes).
The crdS gene (1965 bp) encodes the curdlan synthase (CrdS, 73 kDa) protein. In Agrobacterium, CrdS is an integral inner membrane protein with seven transmembrane (TM) helices, one non-membrane-spanning amphipathic helix and a Nout–Cin disposition. The UDP-Glucose substrate-binding and the catalytic motif for curdlan polymerisation is observed between TM3 and TM4. The process of Curdlan polymerisation mediated by CrdS occurs on the cytoplasmic face of the inner membrane
For our project, we wanted to produce curdlan by E.coli, so this part has been made with a codon optimization of the natural crdS sequence for E.coli.




Curdlan synthase gene CrdS under the control of a stationary phase promoter ( pOsmY ): BBa_K1686033

BBa_K1686033 is a composite part consisting of an Escherichia coli osmY stationary phase promoter (BBa_J45992) and Curdlan synthase gene (BBa_K1686046). Thus, BBa_K1686033 produces curdlan synthase in stationary phase cultures.

See BBa_J45992 for details on the osmY stationary phase promoter.
See BBa_K1686046 for details on the Curdlan synthase gene.




crdA gene with codon optimisation for E. coli: BBa_K1686045




crdC gene with codon optimisation for E. coli: BBa_K1686047




His-Gal promoter BBa_K1686098




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