Protein purification by affinity tags

Jongrak Kittiworakarn g3937506 at STUDENT2.MAHIDOL.AC.TH
Fri Oct 30 19:26:14 EST 1998


> Dear Dr. Chen:

In order to express a protein, the DNA sequence around 5' untranslated region
and also the sequence around starting site of translation may have an
important role.
The fusion protein to the N-terminus of the target protein will make sure that
you have not to make any modification around these sequnces. Also, by
detection of this tag, it will give you some info. about your expression
problem.

Jongrak


> Protein purification by affinity tags
>
> Yu Wai Chen (ywc at mrc-lmb.cam.ac.uk)
> Fri, 30 Oct 1998 16:49:00 +0000
>
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>   ------------------------------------------------------------------------
>
> To: protein-analysis at net.bio.net
>
> From: Yu Wai Chen <ywc at mrc-lmb.cam.ac.uk>
>
> Subject: Protein purification by affinity tags
>
> Date: Fri, 30 Oct 1998 16:49:00 +0000
>
> Hello,
>
> This is a general question about making fusion protein for affinity
> purification. Several protein purification systems offer a choice of N-
> or C- terminal fusion of affinity tags, such as CBD, His6 etc while
> other systems have only N-terminal tags, such as GST, MBP. It is
> obvious that if the tag is at the N-terminus, one can get truncated
> protein products bound onto the affinity column. If the tag is
> C-terminal, only full-length fusion protein will be retented. With such
> consideration, why would so many affinity tags be made to the
> N-terminus? It seems to me that it offers no advantage at all...
>
> Please comment.
>
> --
> ===================================================================
> Yu Wai CHEN, Ph.D. ..................  email: ywc at mrc-lmb.cam.ac.uk
>  Centre for Protein Engineering,              tel: 44-(1223) 402148
>  MRC Centre, Cambridge  CB2 2QH, U.K.         fax: 44-(1223) 402140
>  WWW homepage: http://www.mrc-cpe.cam.ac.uk/people/wai.html
>
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>    * Next message: G. L. Prasad: "Two Postdoctoral Positions available"
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--
-----------------------------------Mr. Jongrak Kittiworakarn
                                   g3937506 at student.mahidol.ac.th

     Inst. of Science and Technology for Research and Development
     Mahidol University (Salaya Campus)
     Salaya, Nokornprathom, THAILAND.
     Tel. 001-66-2-441-9003-7 ext.1277,1278,1279
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