Coding
TSHHantag

Part:BBa_K2404001

Designed by: Ryan Coates   Group: iGEM17_Cardiff_Wales   (2017-09-22)
Revision as of 11:06, 6 October 2017 by Gparry75 (Talk | contribs)


A Thyroid Stimulating Hormone antagonist with His-Tags

This is a coding sequence of a gene that produces a protein that competitively inhibits thyroid stimulating hormone (TSH). This includes a 6xHis-tags to aid protein purification.

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 himans.

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.

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 BamHI site found at 795
    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 827


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