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heterodimer


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#1 Rsm

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Posted 13 May 2009 - 10:43 PM

Hi,

I'd like to express and purify a heterodimeric kinase in bacteria. Is it possible to put both genes into one vector, the second gene with a ribosome binding site in front? The first subunit has then a 6xHis tag, can I purify the whole kinase with this? Would you think that this kinase might work in bacteria, ie. when I further transfect a kinase target, would that be phosphorylated in the bacteria?
Thanks!

Minna
I got soul, but I'm not a soldier

#2 genz

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Posted 15 May 2009 - 10:44 PM

I think you certainly can express two proteins in 1 host using 1 vector. There are vectors called Dual Vector system. I think Merck, Novagen, Invitrogen all have different dual vectors..

#3 Rsm

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Posted 16 May 2009 - 07:30 PM

Thank you!
Unfortunately, our lab budget is a little tight, so I was hoping to circumvent buying a new vector (for example the pETDuet from Novagen) by creating it myself... There is some literature about this, e.g. PMID 19017543, where they expressed two proteins just by integrating a second ribosome entry site. Anyone some experience?
I guess the other questions are not easy to answer, I might need to try and find out myself... :(
Thanks again and good luck to all of you.
Cheers, Minna
I got soul, but I'm not a soldier




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