Protein complexes perform essential functions in cells. Fatty acid synthase in human is a protein complex made of multiple proteins. RuBisCO is another protein complex. They are composed of multiple proteins. What ideas do you have to express those protein complexes in bacteria or yeasts? Or there might be some other solutions like growing tissue cultures containing the complex proteins of interest?
We tried (unsuccessfully) to co-express an in vivo complex in E. coli using the pETDuet system from Novagen.
Hi there,
Cloning genes as an operon in pET plasmid is a solution to be considered (only one gene leading to tagged protein expression for purification purpose). The obvious thing to do is to perform gene synthesis in order to optimize gene sequence for expression in bacteria.
An other option in yeast is the use of pESC plamid serie (4 plasmids available with 4 different auxotrophy markers) in which you can clone two genes per plasmid under inducible GAL promoter.
To express complex active protein, i suggest to use yeast system rather than e.coli system. e.coli system more simple than yeast system but sometimes e.coli makes foreign protein become inactive after expressed. But must be checked for the protein activity. thanks
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