Coding

Part:BBa_K2586020

Designed by: Janek Meißner   Group: iGEM18_Goettingen   (2018-09-13)
Revision as of 14:31, 9 October 2018 by Robertw (Talk | contribs)


AroA*: Resistant aroA-variant in Escherichia coli

This part encodes a resistant variant to the native aroA gene in Escherichia coli. The resistance occurs due to 2 point mutations within the DNA sequence. At position 287 a Guanin was changed to a Cytosin and at position 547 a Guanin was changed to an Adenin.

This part can be used to confer glyphosate resistance to bacteria, as the mutation decreases the interaction of the herbicide with the EPSP-Synthase, which is encoded by the native aroA gene.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 346
    Illegal AgeI site found at 787
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1021


aroA: 3-phosphoshikimate 1-carboxyvinyltransferase

This part encodes for the 3-phosphoshikimate 1-carboxyvinyltransferase. The encoded enzyme is needed for the biosynthesis of essential aromatic amino acids in the shikimate pathway. Three essential aromatic amino acids are products of the pathway: Trp, Phe and Tyr. aroA is expressed in the E.coli wildtype.

aroA encodes the 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase, which is the main target of the herbicide glyphosate and is strongly inhibited through glyphosate application. An amplification of aroE/aroA allows for glyphosate resistance (1), this is used in multiple GM plants to increase the yield and improve farming applications. If exposed to glyphosate, E.coli mutants will make changes in the original sequence of aroA to avoid the lethal effects of the substance.

Fig. 1.Crystal structure of AroA


Locus BSU29750
Isoelectric point 5.34
Molecular weight 39.38 kDa
Protein length 358 aa
Gene length 1074 bp
Function biosynthesis of aromatic amino acids
Product 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase/
chorismate mutase-isozyme 3
Essential no
Synonyms aroG

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Categories
Parameters
None