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
- - - - -

Does KanR gene driven by Kanamycin promoter on a plasmid (e.g., pCR4TOPO) confer

Transfection Neo Kan G418 Cloning

  • Please log in to reply
3 replies to this topic

#1 albertwcheng

albertwcheng

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 06 March 2012 - 10:59 PM

Hi!!

I just noticed that KanR and NeoR used in bacterial selection and mammalian selection are related if not identical. I didn't consider when cloning a puro resistance casette into pCR4 TOPO (which carries the Kanamycin promoter driving KanR gene for bacterial selection). I worry that the KanR gene would be somehow (maybe leakily) expressed from the vector in mammalian cells and it will make that mammalian cells also NeoR (which is undesirable as I only intend to make the transfected cells puro resistance) if transfected with that plasmid. Please help!

Thanks!
Albert

#2 Curtis

Curtis

    Metaller Scientist

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,112 posts
59
Excellent

Posted 16 March 2012 - 07:48 PM

good point, but I wouldn't worry about that. most of pEGFP plasmids have kanamycin.

#3 pDNA

pDNA

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 493 posts
14
Good

Posted 18 March 2012 - 01:08 AM

as long as you have no eukaryotic promoter upstream of your kanamycin/neomycin (whatever) you don't have to worry ...the kan/neo is often driven by the SV40 (like in pcDNA3.1) ...so watch out for this one.

Regards,
p

#4 albertwcheng

albertwcheng

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 18 March 2012 - 11:42 AM

Thanks!





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

©1999-2013 Protocol Online, All rights reserved.