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
- - - - -

RBS site removed in construct. Will there be expression?


  • Please log in to reply
2 replies to this topic

#1 HOYAJM

HOYAJM

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 159 posts
12
Good

Posted 09 February 2012 - 10:37 AM

I inserted a gene of interest into a pET32a vector downstream of the T7 promoter. However, while doing this I also removed the RBS unintentionally. I plan to express the construct in BL21 E. coli to purify the protein of interest. My question is... will there be expression of the protein despite the lack of the RBS? Will the ribosomes be able to translate the mRNA without it? I am doubtful it can, but I am certainly trying to prevent having to make a new construct.


Thanks

Edited by HOYAJM, 09 February 2012 - 10:46 AM.


#2 scolix

scolix

    a student

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 314 posts
5
Neutral

Posted 09 February 2012 - 10:57 AM

I am not sure how it works with prokaryotic expression. But in eukaryotes, there can be reduction in the translation if RBS is lacking.

#3 phage434

phage434

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,418 posts
238
Excellent

Posted 09 February 2012 - 11:25 AM

It might express, but I would not count on it. An experiment will tell you, or you can fix it.




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

©1999-2013 Protocol Online, All rights reserved.