Difference between revisions of "Part:BBa K1216004"

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	<partinfo>BBa_K1216000 short</partinfo>		<partinfo>BBa_K1216000 short</partinfo>
−	''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] ]]	+	''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>]].
−	A form of this protein with added TEV and poly-HIS tags can be found [[Part:BBa_K1216004| here]].	+	A form of this protein without TEV and poly-HIS tags can be found [[Part:BBa_K1216004| here]].
	===Usage and Biology===		===Usage and Biology===
−	β-Glucuronidase is used as a fusion protein marker in higher plants, due to them lacking intrinsic β-Glucuronidase activity[[Part:BBa_K1216000#References|<sup>[2]</sup>]].	+	β-Glucuronidase is used as a fusion protein marker in higher plants, due to them lacking intrinsic β-Glucuronidase activity[[Part:BBa_K1216000#References|<sup>[4]</sup>]].
−	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[[Part:BBa_K1216000#References|<sup>[3]</sup>]].	+	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[[Part:BBa_K1216000#References|<sup>[5]</sup>]].
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	===References===		===References===
	#  [http://ecoliwiki.net/colipedia/index.php/uidA:Gene ecoliwiki]		#  [http://ecoliwiki.net/colipedia/index.php/uidA:Gene ecoliwiki]
		+	# 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]
		+	# [http://homepage.univie.ac.at/nikos.pinotsis/tev_protease.html University of Vienna TEV Protease info]
	# 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/]		# 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/]
	# [http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Bulletin/gusabul.Par.0001.File.tmp/gusabul.pdf Sigma Aldrich]		# [http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Bulletin/gusabul.Par.0001.File.tmp/gusabul.pdf Sigma Aldrich]

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Revision as of 13:02, 6 September 2013

β-Glucuronidase (gusA) from Bacillis Subtilis

gusA (also called uidA^[1]) encodes β-Glucuronidase, an intracellular enzyme that catalyzes the hydrolysis of β-D-glucuronides. 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

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]