Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

expression vector in E. coli


  • Please log in to reply
5 replies to this topic

#1 Alie

Alie

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 05 June 2009 - 03:25 AM

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

#2 Amy831

Amy831

    member

  • Active Members
  • Pip
  • 10 posts
0
Neutral

Posted 05 June 2009 - 09:20 AM

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

#3 labmeiser

labmeiser

    member

  • Active Members
  • Pip
  • 6 posts
0
Neutral

Posted 05 June 2009 - 10:00 AM

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.


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



#4 neuron

neuron

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 187 posts
5
Neutral

Posted 06 June 2009 - 03:32 AM

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

#5 ASITZA

ASITZA

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 07 July 2009 - 01:28 PM

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?

#6 klinmed

klinmed

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 209 posts
1
Neutral

Posted 08 July 2009 - 11:57 PM

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.




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.