Part:BBa_K511601

N-Cadherin-eGFP Cell Adhesion MammoBlock

This part encodes a fusion protein. The first part, N-Cadherin is kmown to be instrumental in intercellular adhesion and tissue integrity. Like all cadherins, N-Cadherin mediates adhesion with five extracellular, calcium-binding repeats and an intercellular domain that makes contacts with the actin cytoskeleton of mammalian cells. The second part is an enhanced green fluorescent protein (eGFP), allowing for expression of both cell adhesion and an identifying marker with a single part.

The construct shown above uses a N-cadherin eGFP fusion protein that allows us to visualize localization of N-cadherin. Using confocal microscopy, we were able to observe membrane localization of our N-cadherin protein as expected. Additionally, we can also observe very bright signal at the cell interfaces, showing clustering of N-cadherin at these interfaces and thus adhesion.

Meanwhile, in our control, we transfected a constitutive GFP construct. We can see diffuse cytoplasmic localization versus membrane localization.

This series of experiments show that our Ncad construct does display expected behavior and provides a promising start to incorporating adhesion into our designs. We observe membrane localization and sticking which is what we were looking for.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 289
  Illegal PstI site found at 1390
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 289
  Illegal PstI site found at 1390
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 922
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 289
  Illegal PstI site found at 1390
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 289
  Illegal PstI site found at 1390
  Illegal NgoMIV site found at 1241
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1276
  Illegal BsaI.rc site found at 1833