I have to express a series of proteins with different molecular weights ranging from 20-36 kDa. I need to express these proteins with pET vectors in E.coli and according to some research articles most of the small mol.wt proteins were expressed in pet15,20,22,23 vectors.
Is there any criteria to choose these vectors other than solubility and disulphide bond formation?
Please provide your suggestions to choose these vectors based on the mol.wt of the protein.
Aside from differences in cloning, each of the different vectors adds different features to the expressed protein. Some of them add perimplasmic localisation tags, some of them add His tags, some add tags to the N-terminal, some to the C-terminal. You can get vectors which add cleavable tags, maltose binding tags, GST tags etc. You need to decide on your purification method (IMAC, MBP etc.) and whether you want to remove the tag. Then you need to decide which end of the protein to add the tag. Finally you can decide on a cloning site, although you can use non restriction enzyme cloning, such as Gibson, so this is less important. It is possible you will need to try different vectors to get good expression of different proteins.
Over the molecular weight range you indicate, all of the pET vectors should work fine in expressing recombinant proteins. Other criteria are much more critical to successful expression, such as the nature of the fusion tag, solubility of the recombinant eukaryotic protein when expressed in a bacterial environment, etc.
In my experience i have used pET vectors to express proteins ranging from 27 kDa to 68 kDa. So you can go ahead and use them in your work. Just be careful in choosing the appropriate tag which you need to use for purification
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