Team:Edinburgh/Cell Display

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Revision as of 09:12, 1 July 2011

An obvious type of bioreactor is an E. coli cell that displays the desired proteins on its outer membrane. This type of display is called cell surface display.

This works by fusing the protein of interest to a carrier protein which is naturally found on the outer membrane.

Notes for standard display

Berkeley 2009 Parts may or may not be helpful: they tried several different carrier proteins. When they tried attaching cellulases, they weren't too successful - of the two quantified cellulases, one worked just as well without the carrier (Cel5b) and the other didn't work (Cel9a, as compared to negative control).

<partinfo>BBa_K265008</partinfo> is the first 211 and last 97 amino acids of ice nucleation protein (INP, coded by [http://www.uniprot.org/uniprot/O30611 inaK] gene) from Pseudomonas syringae. It seems promising as a carrier of enzymes. [http://www.sciencedirect.com/science/article/pii/S016777991000199X Van Bloois et al (2011)] speak highly of INP. Fusions are carried out at the INP C-terminal.

We would be the first team to attempt Ice Nucleation Protein as a carrier. INP has major domains at its N and C terminals, as well as a number of internal repeating domains. There seem to be three strategies for using INP:

• Deleted everything except the N domain; fuse at the new C terminal
• Delete the internal domains; fuse at the C terminal
• Delete nothing; fuse at the C terminal

<partinfo>BBa_K265008</partinfo> should be suitable for the 2nd strategy.

Notes for flagellar display

This may be tricky...

References

• Van Bloois E, Winter RT, Kolmar H, Fraaije MW (2011) [http://www.sciencedirect.com/science/article/pii/S016777991000199X Decorating microbes: surface display of proteins on Escherichia coli]. Trends in Biotechnology 29(2): 79-86 (doi: 10.1016/j.tibtech.2010.11.003).

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