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 a Promoter


  • Please log in to reply
3 replies to this topic

#1 huseler23

huseler23

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 03 January 2011 - 01:30 PM

In order to clone a promoter, does it require ATG start site?

#2 perneseblue

perneseblue

    Unlimited ligation works!

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 578 posts
17
Good

Posted 03 January 2011 - 05:56 PM

The ORF of your gene starts with an ATG codon (most of the time).
Your promoter can start with any sequence. A promoter contains transcription factor binding site, the TATA box and kozak sequence.
May your PCR products be long, your protocols short and your boss on holiday

#3 huseler23

huseler23

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 03 January 2011 - 06:11 PM

The ORF of your gene starts with an ATG codon (most of the time).
Your promoter can start with any sequence. A promoter contains transcription factor binding site, the TATA box and kozak sequence.


For an experimental sample, how far upstream must the promoter be placed?

#4 perneseblue

perneseblue

    Unlimited ligation works!

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 578 posts
17
Good

Posted 03 January 2011 - 07:31 PM

In an experimental system?
As close as you want to. However there are a few papers that report the spacing between promoter and ATG sequence can influence gene expression. The paper reports an optimum distance. Unfortunately I can not remember the details. I usually just stick the promoter immediately adjacent to the gene of interest.

If you are expressing a gene in eukaryotic systems, you need to include an intron for efficient export of the mRNA into the cytoplasm. Some synthetic promoters have the intron included. (ie CAG promoter)
May your PCR products be long, your protocols short and your boss on holiday




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

©1999-2013 Protocol Online, All rights reserved.