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Membrane protein


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#1 manavid

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Posted 17 June 2013 - 08:27 AM

Hello everyone,

I am working on a specific membrane protein. Any suggestions as to which expression host would be best for such proteins (bacteria, yeast) etc.

Thanks.
Manavid

#2 labtastic

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Posted 19 June 2013 - 07:41 AM

It largely depends on the protein you are trying to express. Often (not always) if you're trying to express a eukaryotic protein, insect cells or yeast expression is best. If it's bacterial, then try expressing in e.coli first since it is usually easier that way.

Basically, you will have to try a multitude of expression systems (varying expression host, vector, construct, ortholog, expression time, expression temp, etc) and find the one that works best. If this is a membrane protein that nobody has ever worked with before, be prepared for a marathon project. Figuring out the best expression system can take several months or longer. Once you've found a good expression system, then you need to find detergent(s) that it behaves well in (i.e. it has to be stable, monodisperse, maintains native oligomeric state, active, etc). This can take a long time even with the right equipment.

Hint: you can save yourself a lot of time with these initial screening procedures if you express your membrane protein with a cleavable c-terminal GFP tag.




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