No replies to this topic

[vrindasim23]

Hi all,

I know this is a dumb idea, but I have to ask ....
I have three genes cloned into pET32a+ vector and the protein ( lets say fusion protein-Trx gene +gene of interest) expression in Bl21Plys is good (the protein i get in native conditions and not in inclusion bodies at all) . Purified using His tag columns, that too went fine...now comes the real mess I tried to remove the tag that is the Trx tag using enterokinase ,the protein gets digested with the fusion protein and protein of interest almost running together in PAGE, but not purified using the his tag columns.....
I am running short of time for my thesis work , the real question is has anybody used the trx tagged fusion protein for development of polyclonals in rabbit????
I know its not the real sense to use the fusion protein for polyclonal development, but has anybody tried to do so??

Thank you all.....