Difference between revisions of "Part:BBa K1216004"
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''gusA'' (also called uidA[[Part:BBa_K1216000#References|<sup>[1]</sup>]]) encodes β-Glucuronidase, an intracellular enzyme that catalyzes the hydrolysis of β-D-glucuronides. [[File:Glucuronidase_3d_model.jpg|thumb| 3D representation of the β-Glucuronidase from [http://www.rcsb.org/pdb/explore/explore.do?structureId=3K46 RCSB] ]] The poly-HIS tag can be used for protein purification (IMAC)[[Part:BBa_K1216001#References|<sup>[2]</sup>]]. The TEV tag can then be used to have the TEV protease specifically cleave off the poly-HIS tag from the purified protein [[Part:BBa_K1216001#References|<sup>[3]</sup>]].
 
''gusA'' (also called uidA[[Part:BBa_K1216000#References|<sup>[1]</sup>]]) encodes β-Glucuronidase, an intracellular enzyme that catalyzes the hydrolysis of β-D-glucuronides. [[File:Glucuronidase_3d_model.jpg|thumb| 3D representation of the β-Glucuronidase from [http://www.rcsb.org/pdb/explore/explore.do?structureId=3K46 RCSB] ]] The poly-HIS tag can be used for protein purification (IMAC)[[Part:BBa_K1216001#References|<sup>[2]</sup>]]. The TEV tag can then be used to have the TEV protease specifically cleave off the poly-HIS tag from the purified protein [[Part:BBa_K1216001#References|<sup>[3]</sup>]].

Revision as of 13:05, 6 September 2013

β-Glucuronidase (gusA) from Bacillis Subtilis with TEV and poly-HIS tags

gusA (also called uidA[1]) encodes β-Glucuronidase, an intracellular enzyme that catalyzes the hydrolysis of β-D-glucuronides.
3D representation of the β-Glucuronidase from [http://www.rcsb.org/pdb/explore/explore.do?structureId=3K46 RCSB]
The poly-HIS tag can be used for protein purification (IMAC)[2]. The TEV tag can then be used to have the TEV protease specifically cleave off the poly-HIS tag from the purified protein [3].


A form of this protein without TEV and poly-HIS tags can be found here.

Usage and Biology

β-Glucuronidase is used as a fusion protein marker in higher plants, due to them lacking intrinsic β-Glucuronidase activity[4]. Generally it can be used as reporter enzyme with detection by biochemical activity assays, immunological assays or by histochemical staining of tissue sections or cells[5].


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 538
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



References

  1. [http://ecoliwiki.net/colipedia/index.php/uidA:Gene ecoliwiki]
  2. Loghran ST, "Purification of poly-histidine-tagged proteins.",Methods Mol Biol. 2011;681:311-35. doi: 10.1007/978-1-60761-913-0_17.[http://www.ncbi.nlm.nih.gov/pubmed/20978973]
  3. [http://homepage.univie.ac.at/nikos.pinotsis/tev_protease.html University of Vienna TEV Protease info]
  4. Jefferson A R, "GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.", EMBO J. 1987 December 20; 6(13): 3901–3907 [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC553867/]
  5. [http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Bulletin/gusabul.Par.0001.File.tmp/gusabul.pdf Sigma Aldrich]