Difference between revisions of "Part:BBa K2404000"

Latest revision as of 20:47, 30 October 2017

A Thyroid Stimulating Hormone antagonist without His-Tags

This is a coding sequence of a gene that produces a protein that competitively inhibits thyroid stimulating hormone (TSH).

TSH was first generated and characterised by [http://www.jbc.org/content/276/7/4543.long Fares et al (2001) JBC 276, 4543-4558]. The protein includes two parts, the beta subunit from Thyrotropin (TSH) fused to a common alpha subunit shared by hormone glycoproteins. This is linked by a short CTD linker sequence. This TSH antagonist has been modified to remove glycosylation sites that exist on both subunits. As a result this antagonist binds to the TSH receptor but does not activate it.

TSH has been expressed in a murine expression system and shown to inhibit the auto-immune antibodies that cause in Graves' disease ([http://www.pnas.org/content/113/5/1244.full Saxena et al (2015). PNAS 113, 1244-129]).

We intend to use this TSH protein as a therapeutic treatment to treats hyperthyroidism in humans.

This part has been cloned ito pSB1C3 using BsmBI and conforms to the [http://2016.igem.org/Resources/Plant_Synthetic_Biology/PhytoBricks Phytobrick] standard. It is flanked by BsaI sites together 5' aatg and 3' gctt cloning sequences for subsequent cloning into a level 1 plasmid.

We also generated a version of TSH containing a C-terminal His tag BBa_K2404001

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal PstI site found at 540
Illegal PstI site found at 582
12
INCOMPATIBLE WITH RFC[12]
Illegal PstI site found at 540
Illegal PstI site found at 582
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 607
23
INCOMPATIBLE WITH RFC[23]
Illegal PstI site found at 540
Illegal PstI site found at 582
25
INCOMPATIBLE WITH RFC[25]
Illegal PstI site found at 540
Illegal PstI site found at 582
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1
Illegal BsaI.rc site found at 794

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2404000 short</partinfo>
-This is a coding sequence of a gene that produces a protein that competitively inhibits thyroid stimulating hormone (TSH). It also inhibits auto-immune antibodies that result in Graves' disease. This protein is modified to contain a linker region between two sub units. These sub units are the alpha sub unit which is shared between many human hormones including some sex hormones such as human chorionic gonadotropin (hCG), and the beta sub unit which is specific to TSH. These sub units have been joined in this modified protein by a CTD linker to produce one protein that mimics natural TSH in the body. However, the TSH antagonist lacks glycosylation groups that usually exist on both sub units. This antagonist binds to the TSH receptor but does not activate it as a result. This part would be useful when used as the coding sequence of a gene construct to create a drug that treats hyperthyroidism. This part is contained within the pSB1C3 plasmid.
+This is a coding sequence of a gene that produces a protein that competitively inhibits thyroid stimulating hormone (TSH).
+TSH was first generated and characterised by [http://www.jbc.org/content/276/7/4543.long Fares et al (2001) JBC 276, 4543-4558]. The protein includes two parts, the beta subunit from Thyrotropin (TSH) fused to a common alpha subunit shared by hormone glycoproteins. This is linked by a short CTD linker sequence. This TSH antagonist has been modified to remove glycosylation sites that exist on both subunits. As a result this antagonist binds to the TSH receptor but does not activate it.
+TSH has been expressed in a murine expression system and shown to inhibit the auto-immune antibodies that cause in Graves' disease ([http://www.pnas.org/content/113/5/1244.full Saxena et al (2015). PNAS 113, 1244-129]).
+We intend to use this TSH protein as a therapeutic treatment to treats hyperthyroidism in humans.
+This part has been cloned ito pSB1C3 using BsmBI and conforms to the [http://2016.igem.org/Resources/Plant_Synthetic_Biology/PhytoBricks Phytobrick] standard. It is flanked by BsaI sites together 5' aatg and 3' gctt  cloning sequences for subsequent cloning into a level 1 plasmid.
+We also generated a version of TSH containing a C-terminal His tag [https://parts.igem.org/Part:BBa_K2404001 BBa_K2404001]
 <!-- Add more about the biology of this part here