Part:BBa_K2751014

ALB-mEGFP

Usage and Biology

This part is an improved part of part BBa_S03452, which is a part that contains a EGFP(Enhance Green Fluorescent Protein).

Albumin (ALB) signal peptide can be tagged to protein of interest and allow the protein of interest to be secreted extracellularly. This method has been proved by C. Attallah et al[1]. We improved the part by point-mutating EGFP to changed the Van-Der-Waals force between identical EGFP molecules to decrease EGFP’s propensity to form dimers and by fusing the ALB signal peptide to our mutated EGFP, which becomes monomer EGFP(mEGFP) in order to make the mutated EGFP able to be secreted.

Characterization

We ran electrophoresis to check if the length of the part is correct. Our insert contains ALB sequence and mEGFP sequence. We expect a product of 802bp (834bp along with a biobrick prefix and suffix). Gel photo is shown in the figure below.

reference: 1. A highly efficient modified human serum albumin signal peptide to secrete proteins in cells derived from different mammalian species. (2017, January 10). Retrieved from https://www.sciencedirect.com/science/article/pii/S1046592816305034

Sequence and Features

Assembly Compatibility:

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

  Illegal BamHI site found at 780
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]