Part:BBa_K1061014

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.