5 replies to this topic

[Alie]

Dear everyone

Could you please give me info what is the best expression vector for overexpression recombinant protein in E. coli
Thank you for your help
Alie

[Amy831]

We use pcDNA3.1, which works well for us in E coli.

[labmeiser]

I like the Champion dTOPO directional pET system from Invitrogen.  And expression BL21Stars.  We get about up to 50% of total protein to be our protein.  Only problem is if your protein is toxic, then you have to use a better controlled bacteria, like the BL21 Lys.

Alie, on Jun 5 2009, 06:25 AM, said:

Dear everyone

Could you please give me info what is the best expression vector for overexpression recombinant protein in E. coli
Thank you for your help
Alie

[neuron]

we use pET 21 and pET 28, both are used to express in BL21.

[ASITZA]

Hello everyone!
Could you tell me something more about other procaryotic expression systems, not based on phage T7 elements, such as P BAD, or P TAC? What kind of E.coli stains would I need?

[klinmed]

Amy831, on Jun 5 2009, 07:20 PM, said:

We use pcDNA3.1, which works well for us in E coli.

Don't think you do!
pcDNA3.1 is a mammalian expression vector(CMV prom etc).  It lacks all the upstream regulatory elements needed for expression in E.coli.