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

Mammalian expression vectors with HA, c-myc, FLAG or anyother tag


  • Please log in to reply
3 replies to this topic

#1 Niraj

Niraj

    member

  • Active Members
  • Pip
  • 28 posts
0
Neutral

Posted 12 June 2009 - 04:10 PM

Hi All,
It will be great if somebody can tell me the source for mammalian expression plasmids with tags.
Looking forward to hear from you.
Regards,
Niraj

#2 Dr Teeth

Dr Teeth

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 221 posts
1
Neutral

Posted 15 June 2009 - 07:35 AM

Hi All,
It will be great if somebody can tell me the source for mammalian expression plasmids with tags.
Looking forward to hear from you.
Regards,
Niraj


Try Addgene (http://www.addgene.org/pgvec1). It is a share site that provides plasmids submitted from academic groups in the same way that you request vectors from other labs. You can obtain numerous vectors for about $65 ea. if you sign an MTA.

Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley

#3 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,597 posts
384
Excellent

Posted 22 July 2009 - 04:33 PM

There is also http://plasmid.med.h...erview.jsp#cost for about $45/clone.

Also if you have an empty vector and you want to add a tag, just add the tag to a PCR primer for a target gene and amplify from there. If you add a restriction site on either side of the tag, you should be able to chop it out and re-liagte it into other plasmids too.

#4 fishdoc

fishdoc

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 272 posts
2
Neutral

Posted 22 July 2009 - 04:38 PM

There is also http://plasmid.med.h...erview.jsp#cost for about $45/clone.

Also if you have an empty vector and you want to add a tag, just add the tag to a PCR primer for a target gene and amplify from there. If you add a restriction site on either side of the tag, you should be able to chop it out and re-liagte it into other plasmids too.




I just did this myself using FLAG. Added it to pBluescript, and then added a restriction site in front of the tag, so I can make C-terminal FLAG constructs. Can also insert flanking sequence downstream of the tag if I ever want to express the tagged protein as a single copy in the genome as opposed to plasmid-borne. Went together pretty easily. It required some polymerase, restriction enzymes, and some sequencing. All in all, likely cheaper than what I would've paid (was looking at Sigma for FLAG vectors), and probably more versatile with being able to add flanking sequence behind the tag.


As for places that sell vectors, I remember seeing many at Sigma for FLAG. Almost any supplier of molecular biology supplies probably has a good number of tag vectors to choose from. But as stated above, you'll probably pay at least 5 or 6 times more than if you'd get one from the links above.

Edited by fishdoc, 22 July 2009 - 04:43 PM.





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

©1999-2013 Protocol Online, All rights reserved.