Difference between revisions of "Part:BBa K1061014"

 
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1061014 short</partinfo>
 
<partinfo>BBa_K1061014 short</partinfo>
  
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This is a measurement part. We ligated the 4-copy target of miRNA-122 after eGFP protein. We also get the data of the response of the target toward the dosage of miRNA-122 expression, confirm by fluorence, qRT-PCR and western blot. Using this device, iGEM teams can use it to measure and get enough data for modeling of circuit involve miRNA.
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This is a measurement& improve part. We ligated the 4-copy target of miRNA-122 after eGFP protein. We compare it with the 2-copy target constructed by iGEM teams Heiderberg 2010 (https://parts.igem.org/Part:BBa_K337004). And we verify it use 3 ways: qRT-PCR, western blot and fluorence.Besides, we also get the dosage dependent effect of 2-copy version toward miRNA-122 expression. Using this device, iGEM teams can use it to measure and get enough data for modeling of circuit involve miRNA.
  
 
<!-- Add more about the biology of this part here
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1061014 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1061014 SequenceAndFeatures</partinfo>
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====Functional & quantitive analysis====
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We co-expressed this part with 0.75ug plasmid of miRNA-122 expression vector in bosc, and by several methods, we got the following result:
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[[Image:Target-different_target_number.png|thumb|center|500px|suppression Level is related to target copy number]]
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The suppression effect under fix expression of miRNA-122
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And the data about characterization of 2-copy version will be submitted later.
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Latest revision as of 18:36, 26 September 2013

eGFP->miRNA-122 target

This is a measurement& improve part. We ligated the 4-copy target of miRNA-122 after eGFP protein. We compare it with the 2-copy target constructed by iGEM teams Heiderberg 2010 (https://parts.igem.org/Part:BBa_K337004). And we verify it use 3 ways: qRT-PCR, western blot and fluorence.Besides, we also get the dosage dependent effect of 2-copy version toward miRNA-122 expression. Using this device, iGEM teams can use it to measure and get enough data for modeling of circuit involve miRNA.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 721
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 848
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Functional & quantitive analysis

We co-expressed this part with 0.75ug plasmid of miRNA-122 expression vector in bosc, and by several methods, we got the following result:

suppression Level is related to target copy number

The suppression effect under fix expression of miRNA-122 And the data about characterization of 2-copy version will be submitted later.