GST fusion vector for cloning Pst1 fragment

Randy Haun rhaun at nih.gov
Fri Apr 14 09:22:49 EST 1995


In article <Rosenwasserlab-130495154518 at 198.243.65.220> Rosenwasserlab at njc.org (Calvin and Hobbbes) writes:
>Path: nih-csl!darwin.sura.net!fconvx.ncifcrf.gov!biosci!rutgers!gatech!howland.reston.ans.net!news.sprintlink.net!uunet!boulder!tali.hsc.colorado.edu!NewsWatcher!user
>From: Rosenwasserlab at njc.org (Calvin and Hobbbes)
>Newsgroups: bionet.molbio.methds-reagnts
>Subject: Re: GST fusion vector for cloning Pst1 fragment
>Followup-To: bionet.molbio.methds-reagnts
>Date: Thu, 13 Apr 1995 15:45:17 -0600
>Organization: National Jewish Center for Immunolgy and Respiratory Medicine
>Lines: 24
>Message-ID: <Rosenwasserlab-130495154518 at 198.243.65.220>
>References: <D6uCFn.7uq at news.cis.umn.edu> <1995Apr12.172031.1 at molbiol.ox.ac.uk>
>NNTP-Posting-Host: 198.243.65.220
>In article <1995Apr12.172031.1 at molbiol.ox.ac.uk>, rpgrant at molbiol.ox.ac.uk
>wrote:
>
>> In article <D6uCFn.7uq at news.cis.umn.edu>, shin at cis.umn.edu (Shin Enomoto) writes:
>> > Is there a GST fusion vector suitable for cloning a Pst1 fragment.  We 
>> > are looking for a vector that has NsiI site in the polylinker.
>> > 
>> 
>> 
>> Unlikely, but you could always make your own.  Design the polylinker you want, 
>> get your friendly DNA oligo synthesizer to make it (need fwd and rev strands), 
>> anneal, ligate, presto!
>> 
>> Richard
>
>
>Actually, as stated earlier there is a suitable vector. Pharmingen's
>pAcGHLT-A,B,C vector set. Catalog #21463P. Granted, these are used as
>transfer vectors forBaculoexpression but will work well for this
>application.
>
>Good Luck,
>
>Jon S.

If the Pharmingen vector is not suitable for your purposes, I have a modified
pGEX-2T vector that does not require the use of any specific restriction
enzyme for digesting the target sequence.  It does, however, require PCR
amplification of the target with primers that contain specific 5' extensions.
If you are interested, let me know.

Randy Haun
NIH/NHLBI



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