Part:BBa_K1582010

RFP+inJanus-m Fusion Protein

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 AgeI site found at 555
  Illegal AgeI site found at 667
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 871
  

Usage

Admittedly the separation of proteins is difficult in biological experiments. More often than not, we use chromatography, but it is expensive. Extraction turns to be cheap, however, its low separation rate is really an obstacle.
Here comes the Janus, a kind of amphipathic protein, which is made into a fusion protein with the target and carry it to leave the bulk protein phase. We could use aqueous two-phase systems to make them separated from mixture, according to the property that Janus will direct to the phase of detergent in the system of detergent/polymer. RFP-inJanus fusion protein could be used to verify this purification system.

Biology

inJanus are small secreted fungal proteins which can be found in filamentous fungi, which play a role in a broad range of processes in the growth and development of filamentous fungi. They shows rod-like structure, can be expressed in eukaryotic cells, and are involved in transferring themselves to detergent phase spontaneously. We did some mutations to it, in order to make it expressed in E.coli, and we call it inJanus-m.

Reference

[1]Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, 9751 NN Haren, The Netherlands. Hydrophobins: multipurpose proteins.Annu Rev Microbiol. 2001;55:625-46.