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
0
2 replies to this topic
#1
-
- Active Members
-
- 333 posts
Post Dog
Posted 13 May 2009 - 10:43 PM
I got soul, but I'm not a soldier
#2
-
- Active Members
-
- 14 posts
member
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
-
- Active Members
-
- 333 posts
Post Dog
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
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
