Difference between revisions of "Part:BBa K2404001"

 
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<partinfo>BBa_K2404001 short</partinfo>
 
<partinfo>BBa_K2404001 short</partinfo>
  
This is a coding sequence of a gene that produces a protein that competitively inhibits thyroid stimulating hormone (TSH), but has His-tags for purification. 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.
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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.  
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TSH was first generated and characterised by <a href="http://www.jbc.org/content/276/7/4543.long">Fares et al (2001) JBC 276, 4543-4558</a>. 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.  
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TSH has been expressed in a murine expression system and shown to inhibit the auto-immune antibodies that cause in Graves' disease (<a href="http://www.pnas.org/content/113/5/1244.full">Saxena et al (2015). PNAS 113, 1244-129</a>).
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We intend to use this TSH protein as a therapeutic treatment to treats hyperthyroidism in himans.  
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This part has been cloned ito pSB1C3 using BsmBI and conforms to the Phytobrick standard. It is flanked by BsaI sites together 5' aatg and 3' gctt  cloning sequences for subsequent cloning into a level 1 plasmid.  
  
 
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Revision as of 10:31, 6 October 2017


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 <a href="http://www.jbc.org/content/276/7/4543.long">Fares et al (2001) JBC 276, 4543-4558</a>. 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 (<a href="http://www.pnas.org/content/113/5/1244.full">Saxena et al (2015). PNAS 113, 1244-129</a>).

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 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