Hi,
I would like to express and purify a peptide of 30-40 residue long. I would like to know if I can use pET-31b vector with HIS tag or should I go with pGS-21a with GST tag. It would be helpful if any other expression systems are suggested.
Thank you
Dear Raja Sekhar ,
I Feel pGS-21a with GST tag was better option.,
Before you doing this, make sure about your peptide common purity by using RP-HPLC-LC-MS,
And you first find the property of your peptide based on that only we can able to say perfectly which system is suitable.,
You check the nature also, Hydrophobic peptide means the tag should need to change
For answering this question you need to give details about your peptide, otherwise some common suggestions only possible.,
Good Luck & Regards
R.Selva....
Dear Raja Sekhar ,
I Feel pGS-21a with GST tag was better option.,
Before you doing this, make sure about your peptide common purity by using RP-HPLC-LC-MS,
And you first find the property of your peptide based on that only we can able to say perfectly which system is suitable.,
You check the nature also, Hydrophobic peptide means the tag should need to change
For answering this question you need to give details about your peptide, otherwise some common suggestions only possible.,
Good Luck & Regards
R.Selva....
pGS-21a with GST tag will suitable for you
For purification you try HPLC and then you proceed
My favorite is the HIS tag, but it is really up to your peptide and expression system as mentioned above. It is also important to consider the liker between the tag and peptide and how to remove the tag to get the peptide only. Also, it may be necessary to generate plasmids with the tag at the N or C termini to ensure appropriate expression (His-peptide as well as Peptide-His).
Other tags that maybe useful:
SUMO, TAP, MBP, intein
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