Part:BBa_K5036014

CD63

Part Description

It is a protein which is found on the cell surface and it is a member of the tetraspanin family.that can aid in cell signaling and also regulate movement of vesicles within cells. additionally, it can help in our immune response to various infections.

Usage

We implemented this part in our design to form our loading system for the therapeutic agent to be loaded into exosomes. We have used this part as it is specific for exosome's membrane.

this figure illustrates the structure of CD63 in our loading system. .

Literature characterization

PCR was used to amplify the fragment of the outside membrane of SjCD63. The resulting cDNA was cloned into the pET-28a vector to produce the recombinant protein. The recombinant plasmids were then transformed into E. coli BL21 competent cells. The His-tagged CD63 recombinant protein was expressed in E. coli and purified using a Ni column.

SDS-PAGE of the purified recombinant protein showed a single band of the expected size, indicating that the purified CD63 protein was of high purity . The purified recombinant SjCD63 was further confirmed by mass spectrometry. In summary , a combination of PCR, cloning, expression, and purification techniques was used to produce a high-purity recombinant SjCD63 protein.

Reference

Wang, L., Giri, B. R., Chen, Y., Xia, T., Liu, J., Li, H., ... & Cheng, G. (2018). Molecular characterization, expression profile, and preliminary evaluation of diagnostic potential of CD63 in Schistosoma japonicum. Parasitology research, 117, 3625-3631. Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 307
  Illegal AgeI site found at 159
  
• 1000
  COMPATIBLE WITH RFC[1000]