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
- - - - -

PolyA tail essential?


  • Please log in to reply
3 replies to this topic

#1 moerae

moerae

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 88 posts
0
Neutral

Posted 27 October 2012 - 12:40 PM

Hi,

I'm trying to over-express a protein in zebrafish and I have them in a vector that doesn't have a polyA signal. I will eventually clone it into a vector that has a signal, but I want to just see whether the expression of the protein is correct first. I was going to do an RNA run off and inject into the zebrafish. Will the lack of polyA signal affect the expression of protein?

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,664 posts
396
Excellent

Posted 27 October 2012 - 10:43 PM

Possibly, but not all mRNAs have a poly A tail, so it may be OK.

#3 moerae

moerae

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 88 posts
0
Neutral

Posted 29 October 2012 - 02:12 PM

Thanks for the reply. I'm going to do in vitro transcription using T7/T3 RNA polymerase and the kit I have only adds the 5' cap. Am I right to assume that I'll need a specific kit to add a polyA tail to the end of the RNA transcript?

#4 LabLackey

LabLackey

    member

  • Active Members
  • Pip
  • 29 posts
0
Neutral

Posted 08 November 2012 - 03:33 PM

Usually, yes. I know we have bought a poly(a) polymerase from NEB that works quite well. I think by adenylating your transcript you will see an increase in protein expression. Let us know how it goes.




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

©1999-2013 Protocol Online, All rights reserved.