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
- - - - -

Cloning two genes in the same plasmid with one promotor


  • Please log in to reply
4 replies to this topic

#1 Flo1988

Flo1988

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 08 April 2014 - 08:47 AM

Hello!

I want to coexpress two different genes in E. coli.  Is it possible to clone and coexpress two genes, that are part of the same operon and overlap, in the same MCS of pET26b ?

The idea is to get a polycistronic mRNA and two different proteins: One with a His-Tag and another one without. 

 

-Flo-



#2 phage434

phage434

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,385 posts
228
Excellent

Posted 08 April 2014 - 09:24 AM

This should work.



#3 skepticalguy90

skepticalguy90

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 08 April 2014 - 10:19 AM

Just curious, would you need to make sure to place a shine-dalgarno/rbs before each gene?



#4 phage434

phage434

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,385 posts
228
Excellent

Posted 08 April 2014 - 11:03 AM

That would be a good idea. It certainly would improve expression of the second gene.



#5 Flo1988

Flo1988

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 08 April 2014 - 11:29 PM

I think the first gene already contains a SD Sequence for the translation of the second gene.

Thanks for your replies






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

©1999-2013 Protocol Online, All rights reserved.