Part:BBa_K1582014
BFP+inJanus-m Fusion Protein
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 895
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. BFP-inJanus-m fusion protein could be used to verify this purification system.
Biology
mTagBFP is a monomeric protein with a narrow fluorescence emission spectrum with a maximum at 456 nm. It has a tyrosine-based chromophore, giving it substantially higher brightness, faster chromophore maturation and higher pH stability than blue fluorescent proteins with a histidine in the chromophore.
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.
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