Difference between revisions of "Part:BBa K1720002"
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1720002 short</partinfo>
 
<partinfo>BBa_K1720002 short</partinfo>
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This part is a hypoxia response element.
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This is a part used for coding soluble human guanylate cyclase subunit:Alpha 3.Alpha 3 unit interacts with a beta subunit to form the guanylate cyclase enzyme.
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The hypoxia response element is a minimal cis-regulatory element mediating transactivation by the hypoxia-inducible factor (HIF) in mammalian cells.
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Soluble guanylate cyclases(sGC) are heterodimeric proteins that catalyze the conversion of GTP to 3',5'-cyclic GMP and pyrophosphate. It is important for smooth muscle relaxation in the cardiovascular system. sGC contains an HNOX domain, which serves as a receptor for ligands such as nitric oxide, oxygen and nitrovasodilator drugs.There are several different isoforms of sGC subunit ,but the cardiovascular system abundant with alpha 3 unit and beta 3 unit.  
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This element with a GFP reporter was then transfected into HEK293 cells.We use Vivo green fluorescence signal was observed under fluorescence microscope,  
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Before using this part please add a prompter, in our project we add an EF1A promoter to this part.Since EF1A contains Pstl site,we did not add it to this part.
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This part with a GFP reporter was then transfected into HEK293 cells by lentiviral Vector and in vivo green fluorescence signal was observed under fluorescence microscope, it meant that this part was transfected into HEK293 cells successfully.Then we used qRT-PCR to observe transcriptional level of alpha 3 unit.Finally we detected the cGMP concentration with Elisa Kit.
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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 08:04, 26 August 2015

Hypoxia-induced promotor This part is a hypoxia response element. The hypoxia response element is a minimal cis-regulatory element mediating transactivation by the hypoxia-inducible factor (HIF) in mammalian cells. This element with a GFP reporter was then transfected into HEK293 cells.We use Vivo green fluorescence signal was observed under fluorescence microscope,

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 41
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 41
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 41
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 41
  • 1000
    COMPATIBLE WITH RFC[1000]