Device

Part:BBa_K812013

Designed by: Cyrille Pauthenier   Group: iGEM12_Evry   (2012-09-18)
Revision as of 23:37, 26 September 2012 by TriCer (Talk | contribs)

GFP-AID OsTirI polysistronic system for auxin detection in tadpole

This part is the combination of the BBa_K812010(GFP-AID) and BBa_K812012 (OsTirI) for co-expression in frogs. It is impossible to create polysistronic mRNA in tadpole. In order to overcome the problem, we fused the proteins with a self-cleavable peptide, Pep2A. The protein is transcribed as a single protein and the the peptide cleave itself or with the help of an endogeneous protease giving the two protein, colocalized in the same cell.

This system is a modified version of a pattented device by Kanemaki Masato, Kakimoto Tatsuo, Nishimura Kohei, Takisawa Haruhiko and Fukagawa Tatsuo for yeast and mammalian cells use (http://www.freepatentsonline.com/y2012/0115232.html). However the pattent does not cover the use for oviparian such as frogs and chicken.

This part can be micro-injected using pSC2+ ( BBa_K812000) or related plasmid as a backbone.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1990
    Illegal NotI site found at 1844
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3205
    Illegal XhoI site found at 1913
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1612
    Illegal NgoMIV site found at 1721
    Illegal AgeI site found at 1021
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1406
    Illegal BsaI site found at 1636


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Categories
Parameters
None