Stable Clone establishment using Hygromycin or Puromycin vectors - (Jan/26/2007 )

Hi,

I've established stable ECV304 clones using Neomycin and want to stably coexpress a second gene in those cells. Which vector should I use, which antibioticum?
What about the clontech pTK-Hyg Vector? Or invitrogen pIND-Hyg? What are you using? I would prefer a vector with a strong promotor, maybe CMV and with many unique restriction sites in the MCS.
Thanx for your suggestions.

Julian

-SDT52-

We have a zeocin vector (Invitrogen) with CMV promoter. worked nicely to make stable clones expressing more than 1 gene.

-scolix-

QUOTE (scolix @ Jan 26 2007, 03:43 PM)

We have a zeocin vector (Invitrogen) with CMV promoter. worked nicely to make stable clones expressing more than 1 gene.

may I ask you which one (article number)?
Is it just necessary to insert my gene of interest at the MCS starting with ATG, regardless of the distance to CMV promoter or do I have to think of something else?

best regards
Julian

-SDT52-

QUOTE (SDT52 @ Jan 26 2007, 08:53 AM)

QUOTE (scolix @ Jan 26 2007, 03:43 PM)

We have a zeocin vector (Invitrogen) with CMV promoter. worked nicely to make stable clones expressing more than 1 gene.

i am sorry, I dont have any catalog number. Its a pcDNA3.1+ vector with zeocin. You could clone your gene anywhere in the MCS. Shouldnt be a problem.

Good Luck !!!

-scolix-