Coding

Part:BBa_K5036013

Designed by: Emad hamdy Matter   Group: iGEM24_AFCM-Egypt   (2024-09-12)


MCP

Part Description

This MS2 coat protein(MCP) exhibits a strong affinity for a specific stem-loop structure (MS2). This binding ability make it a valuable tool for studying RNA localization, purification, and interactions. It can be used for RNA Visualization By fusing it to fluorescent protein which in turn can be used to study RNA-protein interactions.

Usage

We incorporated this component in two applications. First, it served as a bridge between MS2 and an MMP9 nanobody in our switch. When MMP9 levels increased, it bound to both nanobodies, initiating mRNA circulation and YAP-1 protein production. Second, it was used in our loading system to bind to MS2 and facilitate the delivery of our cargo into exosomes.

this figure illustrates the structure of MCP and its two applications in our design.

Dry lab Characterization

We measured the effect of the nanobodies on the MCP-MS2 binding stability, so we measured MCP-MS2 before and after binding to nanobody1

MCP-MS2 binding stability

Alignment Plot

3D structure of MCP binded to MS2-Aptamers

this figure show That the alignment plot scores high diagonal intensity which indicates the similarity between our structures and the experimental one .


MS2-MCP-NB1 binding stability

Alignment Plot

3D structure of MCP binded to MS2-Aptamers and NB1

this figure show That The alignment plot scores higher diagonal intensity than MCB-MS2 plot which indicates higher protein stability in the presence of NB1 .

Literature characterization

This research involved purifying MS2 coat protein that was first produced using a plasmid. Gel electrophoresis was then used to assess the cleanliness of the isolated coat protein.

This figure indicates the purity of MS2 coat protein at each stage of the purification procedure using gel electrophoresis.

Reference

Peabody, D. S. (1990). Translational repression by bacteriophage MS2 coat protein expressed from a plasmid. A system for genetic analysis of a protein-RNA interaction. Journal of Biological Chemistry, 265(10), 5684-5689.‏

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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