Difference between revisions of "Part:BBa K732000"

 
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<partinfo>BBa_K732000 short</partinfo>
 
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This sequence can be used to express hCG for any downstream uses, and also as an output signal since it triggers pregnancy tests at low concentrations when properly folded.
 
This sequence can be used to express hCG for any downstream uses, and also as an output signal since it triggers pregnancy tests at low concentrations when properly folded.
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'''Proof-of-principle'''
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''E. coli'' were transformed with pET30a vector containing His-tagged hCG-β under the control of a strong promoter and the lac operator. In the basal state, lacI repressor binds to the operator and blocks transcription of hCG-β. Cells were induced with IPTG, which causes a conformational change in the repressor that unblocks transcription.
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Folded protein mixture was applied to a pregnancy test kit, and the control band appeared. The test strip was allowed to sit overnight, and by the next day the test band had appeared, demonstrating detection of hCG-β.
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Test band was not observed when extract was applied without folding solution.
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[[Image:Virginia_2012_Aug_WT-HCG_Preg_Test.jpg]]
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 04:27, 10 October 2012

Human Chorionic Gonadotropin, Beta Subunit (hCG-β)

Human chorionic gonadotropin (hCG) is a hormone produced during pregnancy. Its beta subunit (hCG-β) contains epitopes recognized by pregnancy tests. The full hCG dimer is also used as an ovulation inducer and can stimulate the production of testosterone in men.

This sequence can be used to express hCG for any downstream uses, and also as an output signal since it triggers pregnancy tests at low concentrations when properly folded.

Proof-of-principle

E. coli were transformed with pET30a vector containing His-tagged hCG-β under the control of a strong promoter and the lac operator. In the basal state, lacI repressor binds to the operator and blocks transcription of hCG-β. Cells were induced with IPTG, which causes a conformational change in the repressor that unblocks transcription.

Folded protein mixture was applied to a pregnancy test kit, and the control band appeared. The test strip was allowed to sit overnight, and by the next day the test band had appeared, demonstrating detection of hCG-β.

Test band was not observed when extract was applied without folding solution.

Virginia 2012 Aug WT-HCG Preg Test.jpg


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 199
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 199
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 199
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 199
    Illegal NgoMIV site found at 169
    Illegal NgoMIV site found at 213
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 303