Coding

Part:BBa_K2929003

Designed by: James Hammond   Group: iGEM19_St_Andrews   (2019-10-11)
Revision as of 20:02, 19 October 2019 by Zsoltsemperger (Talk | contribs)


Beta-glucosidase A from Thermatoga maritima (bglA)

bglA gene which encodes the Beta-glucosidase A from thermophile Thermotoga maritima. Translated protein has a mass of 52kDa. The protein contains a triple salt bridge motif which makes it highly thermostable.


Usage and Biology

The Thermatoga maritima Beta-glucosidase A enzyme expresses well under the T7 promoter in a pET28a, even without IPTG induction, due to a leaky promoter. The protein is mostly soluble based on small scale expression tests and runs at ~55 kDa on an SDS-Page gel.

Figure 1. Soluble expression for the wild-type protein, the pET28a vector was known to have a leaky promoter so the bands at ~55kDa (expected mass 52kDa) indicate expression, as they are absent in lanes used for mOrange and Antibody expression (lanes 2-10).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 998
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 927
    Illegal SapI site found at 379


[edit]
Categories
//cds/enzyme
//chassis/prokaryote/ecoli
//function/degradation/cellulose
extremophiles
Parameters
hill_coeficient
protein