Composite

Part:BBa_K1765000

Designed by: Kinuko Ueno   Group: iGEM15_Tokyo-NoKoGen   (2015-09-06)

Pconst(low)-G3DH-T0 terminator-Pconst(low)-OtsB-OtsA-T0 terminator

Pconst(low)-G3DH-T0 terminator-Pconst(low)-OtsB-OtsA-T0 terminator is a composite part. It codes G3DH and OtsA, OtsB together under constitutive promoters. This part expresses the enzyme, G3DH which is essential for inhibition of termite's metabolic pathway. Also, this part codes two enzymes, OtsA, OtsB which supports the function of G3DH.


Applications of BBa_K1765000

BBa_K1765000 is coding three enzymes, G3DH, OtsA and OtsB. Two enzymes, OtsA and OtsB can convert glucose into trehalose. On the other hand, G3DH is can convert trehalose into 3'3- diketo trehalose. In the iGEM project of Tokyo-Nokogen 2015, we focused on the 3'3-diketo trehalose. We think 3'3-diketo trehalose can inhibit the activity of the enzyme, trehalase, which is essential for insects, such as termites.
NKG3DH.jpg

We checked the expression of G3DH by SDS=Page. Following figure shows the result of SDS-Page. From this result, this biobrick parts can produce the three enzymes, G3DH and OtsA, OtsB correctly. Also, we checked the function of OtsA, OtsB. We use glucose as the substrate of these two enzymes. We checked whether OtsA, OtsB can produce trehalose from glucose by TLC. Following figure shows the result of the TLC. The result shows that the OtsA, OtsB expressed form this biobrick can convert glucose and produce trehalose.

SDS-Page.jpg

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 2409
    Illegal NheI site found at 2899
    Illegal NheI site found at 2922
    Illegal NotI site found at 390
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2715
    Illegal BglII site found at 3426
    Illegal BamHI site found at 4181
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 533
    Illegal NgoMIV site found at 630
    Illegal NgoMIV site found at 1846
    Illegal NgoMIV site found at 2142
    Illegal NgoMIV site found at 2695
    Illegal NgoMIV site found at 2760
    Illegal AgeI site found at 812
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1903
    Illegal SapI site found at 2007


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