How to induce expression in mammalian cells with the pcDNA/V5/GW vector ? - (Jul/23/2008 )

I am using the pcDNA/V5/GW vector from Invitrogen. When I expressed in E. coli, I induced with IPTG, but I am not sure how to induce expression in cells which have this vector. Thanks. You can find a map of the vector here: http://tools.invitrogen.com/content/sfs/ve...gwdtopo_map.pdf

-A148-

I don't think you need to do anything; just transfect and grow. The CMV promoter is what induces transcription in most cells types (I've only worked with limited, immortalized mammalian cells).

-Judes-

QUOTE (Judes @ Jul 23 2008, 05:23 AM)

I don't think you need to do anything; just transfect and grow. The CMV promoter is what induces transcription in most cells types (I've only worked with limited, immortalized mammalian cells).

Thanks! I thought they would just start expressing the protein, but I wasn't sure. That means that I need to find a new vector. I want to be able to control the expression. Thanks for your help!

-A148-

QUOTE (A148 @ Jul 23 2008, 09:20 AM)

QUOTE (Judes @ Jul 23 2008, 05:23 AM)

I don't think you need to do anything; just transfect and grow. The CMV promoter is what induces transcription in most cells types (I've only worked with limited, immortalized mammalian cells).

Thanks! I thought they would just start expressing the protein, but I wasn't sure. That means that I need to find a new vector. I want to be able to control the expression. Thanks for your help!

If you want to control expression, you will need an inducible vector. Invitrogen has some tetracycline inducible ones.

-scolix-