GFP truncation - summary

[Tomas Drgon]

Thanx everybody for suggestions. Here's the summary:



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In article <tomasd-0602961225570001 at fp5-9n.domain>,
tomasd at bdg10.niddk.nih.gov (Tomas Drgon) wrote:

> Hi.
> Does somebody know what happens to the GFP if I chop off C-terminal say 30
> amino acids (BstBI site)? Will it still shine?
> 
> Tomas Drgon
> 
> -- 
> Signature under construction...
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    Feb 96 00:24 EST
Date: Wed, 7 Feb 1996 00:24:34 -0500 (EST)
From: Wei Hu <bz182 at freenet.toronto.on.ca>
Subject: Re: GFP

the answer is 'no' as i tried it.

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From: Daniel Gonzalez <meton at rci.rutgers.edu>

Subject: GFP truncation at Bst BI site.

Hello,

A truncation of 30 terminal AAs will not allow the protein to fold properly,
hence it will not "shine".  These are experiments our lab has done over 3
years ago and has not published, I hope that answers your question.

---Daniel Gonzalez
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From: Peter Wang <plw at mrc-lmb.cam.ac.uk>
Organization: Centre for Protein Engineering

Subject: Re: GFP truncation

Tomas:

Apparently you cannot shorten GFP significantly (1 residue at the amino-end 
and <10-15 at the carboxy-end, see reference below) and still have 
fluorescence.

There is a newsgroup just about fluorescent proteins, especially GFP:
     BIONET.MOLBIO.PROTEINS.FLUORESCENT
and there was a recent similar question and reply, which I've clipped in 
below.

krasel at wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) wrote:
>Stefan Wieland (swieland at sun2.ruf.uni-freiburg.de) wrote:
>> We are looking for the shortest,but still functional GFP protein.
>> Does someone know, whether one can truncate GFP without loosing activity?
>
>Not much. Check out
>
>@article{cubitt:95,
>        author  = {Andrew B. Cubitt and Roger Heim and Stephen R. Adams
>                   and Aileen E. Boyd and Larry A. Gross and Roger Y. Tsien},
>        title   = {Understanding, improving and using green fluorescent
>                   proteins.},
>        journal = TIBS,
>        volume  = 20,
>        pages   = {448--455},
>        year    = 1995
>}

---------------------------------------------------------
Peter Wang, M.D., Ph.D.
MRC Centre for Protein Engineering,
Hills Road, Cambridge, CB2 2QH, England

Tel (01223) 402104  (international calls +44-1223-402104)
Fax (01223) 402140  (     "          "   +44-1223-402140)
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