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
- - - - -

How to get pure protein of a gene

pure protein

  • Please log in to reply
3 replies to this topic

#1 saimasbs

saimasbs

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 26 April 2012 - 09:02 AM

i m working on protein expression . i hav only available option to cut my gene with Nde1 or Nco1 to express it in pet system but these enzymes r cutter of my gene also. plz guide me how can i get pure protein of my gene .

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,797 posts
406
Excellent

Posted 26 April 2012 - 01:24 PM

PCR out the gene using primers with appropriate restriction sites, then clone the PCR product into your vector of choice.

#3 saimasbs

saimasbs

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 03 May 2012 - 07:17 AM

thanks bob1 for replying.i hav cloned my gene in PTZ57R/T vector.now i hav to express and purify protein .my gene size is about 3kb and i hav introduced ECORI AND HINDIII sites in it for clonning it in pet21a for expression. but there will be some extra protein with my gene product so how can i will i purify it?

#4 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,797 posts
406
Excellent

Posted 03 May 2012 - 01:04 PM

How will there be extra protein? Your restriction sites should be outside the start and stop codons...





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

©1999-2013 Protocol Online, All rights reserved.