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From owner-fluorescent@net.bio.net Sat May 01 23:00:00 1999
Path: biosci!agate!newsfeed.berkeley.edu!newsfeed.cwix.com!204.127.161.3!wn3feed!worldnet.att.net!wnmaster2!not-for-mail
From: Chris Tharrington <briarley@worldnet.att.net>
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Re: pEGFP-N1?
Date: Sun, 02 May 1999 10:49:10 -0400
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Hi

    Amp and Kan are NOT interchangable- you must use kanamycin.  That
stupid vector drove me crazy for about a week until I realized it had KanR
not AmpR.  Another word of advice - beware of Dam methylated sites -
especially highly attractive unique Xba1 sites - in many Clontech
vectors.  You might want to re-transform into a Dam- strain right from the
get-go.

Dr Lester Davids wrote:

> I am using the pEGFP-N1 vector and really struggling to transform high
> competent JM109's. Has anyone gotten away with using ampicillin as a
> selective antibiotic for this vector or must one use kanamycin?
>
> Also, has anyone had success with using more than 1-10ng DNA of this
> vector in the transformation reaction?
>
> Thanx


From owner-fluorescent@net.bio.net Sat May 01 23:00:00 1999
Path: biosci!LIS.CH!beat.ludin
From: beat.ludin@LIS.CH (Beat Ludin)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Re: green flourocent proteins
Date: 2 May 1999 03:42:59 -0700
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>Hi, i'm a master student in agriculture and i would appreciate if you =
>can help me to understand how a green flourocent proteins can mark =
>mitochondria for a flourocent microscop.

Sure. all you need to do is add a mitochondrial targeting sequence 
that will make the cell import the GFP into mitochondria. There is 
some literature around. As far as I remember, the first one was

Rizzuto et al. Curr Biol 1995 5:635-42

or in plants

Kohler et al. Plant J 1997 11:613-21

Clontech also sells a mitochondrially-targeted GFP, see 
Clontechniques Oct98 and Jan99. This is the vector published in the 
first reference, I believe.

BTW, there are also excellent fluorescent stains specific for (live) 
mitochondria, such as rhodamine 123 and Molecular Probes' excellent 
MitoTracker series.

Cheers,

Beat


LIFE IMAGING SERVICES
               - visit our new web site at www.lis.ch
+-----------------------------------------------------------
| Dr. B. Ludin
| Life Imaging Services     fon ++41 (0)79 235 7154
| Muehletalweg 22           fax ++41 (0)86 062 296 3160 NEW!
| CH-4600 Olten             beat.ludin@lis.ch
| Switzerland               http://www.lis.ch

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From owner-fluorescent@net.bio.net Sun May 02 23:00:00 1999
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From: squillae@STJOHNS.EDU (Squillante Emilio)
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subscribe squillae@stjohns.edu

Emilio Squillante III, Ph.D.
Room 105 St. Albert's Hall
St. John's University - PAS
8000 Utopia Parkway
Jamaica, NY 11439
(718) 990-5020 fax 6316
Office hours T&R 9:30-11; W 8-10 



From owner-fluorescent@net.bio.net Sun May 02 23:00:00 1999
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From: meton@rci.rutgers.edu (Daniel Gonzalez)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Second International Green Fluorescent Protein Symposium
Date: 3 May 1999 16:38:45 -0400
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Summary: Second International Green Fluorescent Protein Symposium
Keywords: Symposium Green Fluorescent Protein GFP
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                      GFP2 Announcements:
The Webpage for the symposium has been updated and now has sessions and 
dates listed, see http://www.rci.rutgers.edu/~meton/GFP2.html.  The cut-off 
for making reservations at the Town & Country (1-800-772-8527) is the 1st 
of May, after that date the special rate for attendees of our meeting may 
not be honoured, as the hotel will try to book the rooms reserved for our 
meeting to other travellers.
The late registration deadline has been moved back to May 7th, for anyone 
registering from the internet off of our webpage, if anyone has any 
questions with this please contact Bill Ward (732-932-9071 ext. 216).
Please consult the following table for an idea of our registration rates.
Please note the discount for people from Rutgers, Scripps and UCSD.

     REGISTRATION RATES FOR UCSD, SCRIPPS, AND RUTGERS PERSONNEL
     -----------------------------------------------------------
     STUDENTS (Full time students and postdocs)   $150.00
     FACULTY AND STAFF                            $297.50

     Other registration rates after May 7 will be:

     REGULAR REGISTRATION RATES
     -----------------------------------------------------------
     STUDENTS (Full time students and postdocs)   $300.00
     NON-PROFIT ORGANIZATIONS                     $595.00
     CORPORATIONS AND FOR-PROFIT ORGANIZATIONS    $895.00

Note:  If you have registered and have not been confirmed withing 3 working 
days, contact us immediately so that we can expedite your registration.

Those speakers who have not sent me an abstract and those people 
who wish to display posters at the meeting should go to the Symposium 
Website; http://www.rci.rutgers.edu/~meton/GFP2.html and go to the abstract 
page linked from that site; http://www.rci.rutgers.edu/~meton/Abstract.html
to get the details for that.  The deadline for which is May 7th.  

Daniel Gonzalez



From owner-fluorescent@net.bio.net Sun May 02 23:00:00 1999
Path: biosci!SUN.BQ.UB.ES!ferrer
From: ferrer@SUN.BQ.UB.ES ("Joan C. Ferrer")
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: (none)
Date: 3 May 1999 08:25:25 -0700
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Dear all,

Does anyone know of a vector suited for the expression in yeast of proteins
fused to GFP? This is, a vector with a promotor that works in yeast and a 
decent MCS for C-terminal fusion of a protein to GFP.
Thanks


From owner-fluorescent@net.bio.net Mon May 03 23:00:00 1999
Path: biosci!HOTMAIL.COM!alexwebb
From: alexwebb@HOTMAIL.COM ("alex webb")
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: LKB1251 Software Help
Date: 4 May 1999 06:07:01 -0700
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I have a LKB1251 luminometer. It is a rather ancient piece of equipment, but 
it does meet my needs until I can get some sophisticated hardware. 
Unfortunately, the hard disk of the computer died and I no longer have the 
software that can run the LKB1251. Wallac no longer support this piece of 
equipment and can not supply me with the software. Is there anybody out 
there who could be kind enough to let me have a copy of the software for 
this luminometer?

Please email on aarw2@cam.ac.uk.

Dr Alex Webb
Department of Plant Sciences
University of Cambridge
Downing Site
Downing Street
CAMBRIDGE
CB2 3EA
UK

Tel:- 01223 333948
Lab:-01223 333928
Fax:- 01223 333953
Email:- aarw2@cam.ac.uk


______________________________________________________
Get Your Private, Free Email at http://www.hotmail.com

From owner-fluorescent@net.bio.net Mon May 03 23:00:00 1999
Path: biosci!AELIAN.GEOG.NOTTINGHAM.AC.UK!houwuehae79
From: houwuehae79@AELIAN.GEOG.NOTTINGHAM.AC.UK (rwegyua)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Quick Money.
Date: 4 May 1999 07:05:07 -0700
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Hello.  This is the answer to :

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From owner-fluorescent@net.bio.net Mon May 03 23:00:00 1999
Path: biosci!utkux.utcc.utk.edu!vonarnim
From: vonarnim@utkux.utcc.utk.edu
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: EGFPs in plants
Date: 4 May 1999 13:48:43 -0700
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Hi: I'd like to find out whether Clontech's EGFPs, and especially *EYFP*
have been tested for expression and fluorescence in plant cells. What were
the results, and in which cell type were the results obtained?  Thanks,

Albrecht


Albrecht von Arnim

   Assistant Professor
   Dept. of Botany
   University of Tennessee
   Knoxville, TN 37996-1100
   vonarnim@utk.edu
   Phone: (423) 974-6206
   Fax:   (423) 974-0978



From owner-fluorescent@net.bio.net Mon May 03 23:00:00 1999
Path: biosci!HOTMAIL.COM!carldahlb
From: carldahlb@HOTMAIL.COM ("Carl Dahlberg")
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Alternatives to FITC-labeling of a FICOLL
Date: 4 May 1999 23:38:20 -0700
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Hi
I am looking for an alternative to FITC.  One that is not poisonous and
thereby possible to give to patients. We have been detecting renal clearance
using FITC-Ficoll. I need one that is possible to bond to a Ficoll molecule.
Ficoll is a big carbohydrate that actually makes the FITC quite stable and
probably not dangerous but it would be a lot handier to use something that
already has been used.

Carl Dahlberg  Department of Physiologi University of Gothenburg Sweden

reply to carldahlb@hotmail.com


From owner-fluorescent@net.bio.net Tue May 04 23:00:00 1999
Path: biosci!U.WASHINGTON.EDU!moser
From: moser@U.WASHINGTON.EDU ("'Mike' Michael J. Moser")
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Re: (none)
Date: 5 May 1999 08:06:59 -0700
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Joan

Tim Stearns at Stanford University put together a series of GFP fusion
vectors enabling expression of either C- or N-terminal GFP fusion proteins
driven by the GAL1 promoter.  I am assuming you mean S. cerevisiae when
you say "yeast".

Happy GFPing

Mike Moser, Ph.D.                                     Tel: 206-543-6585
UW Department of Pathology                            FAX: 206-543-3967
Box 357705                                       moser@u.washington.edu
Seattle, WA  98195                 http://weber.u.washington.edu/~moser

On 3 May 1999, Joan C. Ferrer wrote:

> Dear all,
> 
> Does anyone know of a vector suited for the expression in yeast of proteins
> fused to GFP? This is, a vector with a promotor that works in yeast and a 
> decent MCS for C-terminal fusion of a protein to GFP.
> Thanks
> 
> 
> 


From owner-fluorescent@net.bio.net Tue May 04 23:00:00 1999
Path: biosci!U.WASHINGTON.EDU!moser
From: moser@U.WASHINGTON.EDU ("'Mike' Michael J. Moser")
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Re: Staining dead yeast for FACS
Date: 5 May 1999 08:02:36 -0700
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Tim

The classic live/dead stain in yeast is methylene blue, but I'm not sure
how well it works for flow.  We use propidium iodide (PI) for mammalian
cells.  Might work for you.  Kill some cells with heat or formaldehyde and
try staining.

Happy GFPing

Mike Moser, Ph.D.                                     Tel: 206-543-6585
UW Department of Pathology                            FAX: 206-543-3967
Box 357705                                       moser@u.washington.edu
Seattle, WA  98195                 http://weber.u.washington.edu/~moser

On Wed, 5 May 1999, T.J. Young wrote:

> Dear All,
> I'm using FACS to look at pombe expressing GFP linked to various versions 
> of my protein.  I'm constantly getting a population of low fluorescent 
> cells which I think are probably dead.  Is there a staing I can use to gate 
> out these cells?
> Thanks in advance,
> Tim Young ;-)
> 
> 


From owner-fluorescent@net.bio.net Tue May 04 23:00:00 1999
Newsgroups: bionet.molbio.proteins.fluorescent
Path: biosci!agate!newsfeed.berkeley.edu!dispose.news.demon.net!demon!newspeer.clara.net!news.clara.net!nntp.news.xara.net!xara.net!server5.netnews.ja.net!leeds.ac.uk!news
From: BMBTJY@leeds.ac.uk (T.J. Young)
Subject: Staining dead yeast for FACS
Message-ID: <7got8p$8fk_001@leeds.ac.uk>
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Dear All,
I'm using FACS to look at pombe expressing GFP linked to various versions 
of my protein.  I'm constantly getting a population of low fluorescent 
cells which I think are probably dead.  Is there a staing I can use to gate 
out these cells?
Thanks in advance,
Tim Young ;-)

From owner-fluorescent@net.bio.net Tue May 04 23:00:00 1999
Path: biosci!netsite.com.br!aquarium
From: aquarium@netsite.com.br ("Aquarium Club")
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: urgent!!!
Date: 5 May 1999 18:32:30 -0700
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Please, I'm student the biology and I have seminary. I will like some photo
of expression of plant the GFP and GUS and LUC of the apresentacion the
seminary in 10 the may...
                    tank you...
                                     Simone
e-mail: tepicchi@netsite.com.br or
           scpicchi@fcav.unesp.br

From owner-fluorescent@net.bio.net Wed May 05 23:00:00 1999
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From: Kasper Dreyer <kasper@imsb.au.dk>
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Info on length of GFP linker ????
Date: Thu, 06 May 1999 09:34:03 +0200
Organization: Aarhus Universitet
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Hi

I'd be happy to recieve any information on how long (amino acids) the "linker"
should be between GFP and the protein it is fused to.

Yours Sincerely,

Kasper Dreyer
University of Aarhus, Denmark


From owner-fluorescent@net.bio.net Wed May 05 23:00:00 1999
Path: biosci!U.WASHINGTON.EDU!moser
From: moser@U.WASHINGTON.EDU ("'Mike' Michael J. Moser")
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Re: Info on length of GFP linker ????
Date: 6 May 1999 10:31:29 -0700
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Kasper,

I used 10AA in yeast and most fusions complemented deletion of essential
genes

Mm

Mike Moser, Ph.D.                                     Tel: 206-543-6585
UW Department of Pathology                            FAX: 206-543-3967
Box 357705                                       moser@u.washington.edu
Seattle, WA  98195                 http://weber.u.washington.edu/~moser

On Thu, 6 May 1999, Kasper Dreyer wrote:

> 
> Hi
> 
> I'd be happy to recieve any information on how long (amino acids) the "linker"
> should be between GFP and the protein it is fused to.
> 
> Yours Sincerely,
> 
> Kasper Dreyer
> University of Aarhus, Denmark
> 
> 
> 


From owner-fluorescent@net.bio.net Thu May 06 23:00:00 1999
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From: Mike Challen <mike.challen@hri.ac.uk.nospamthanks>
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: eGFP vs sGFP
Date: Fri, 07 May 1999 09:40:29 +0100
Organization: HRI
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Does anybody know please, what is sGFP?
Is it the same as eGFP, and if not, where can I find the s sequence?  Is
there a reference etc. etc?  My efforts to find sGFP in EMBL have not
been successful.

Thanks

Mike


From owner-fluorescent@net.bio.net Sun May 09 23:00:00 1999
Path: biosci!FNS1.U-SHIZUOKA-KEN.AC.JP!niwa
From: niwa@FNS1.U-SHIZUOKA-KEN.AC.JP (Yasuo Niwa)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Re: eGFP vs sGFP
Date: 10 May 1999 00:42:47 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
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Please refer to
Haas, J., Park, E.-C., and Seed, B. (1996) Codon usage limitation in the
expression of HIV-1 envelope glycoprotein. Current Biology 6, 315-324.

Chiu, W-I., Niwa, Y., Zeng, W., Hirano, T., Kobayashi, H., and Sheen, J. (1996)
Engineered GFP as a vital reporter in plants. Current Biology 6, 325-330.

Yasuo Niwa

At 5:40 PM 99.5.7, Mike Challen wrote:
> Does anybody know please, what is sGFP?
> Is it the same as eGFP, and if not, where can I find the s sequence?  Is
> there a reference etc. etc?  My efforts to find sGFP in EMBL have not
> been successful.
>
> Thanks
>
> Mike



From owner-fluorescent@net.bio.net Mon May 10 23:00:00 1999
Path: biosci!tag.csiro.au!adrian.elliott
From: adrian.elliott@tag.csiro.au ("Adrian Elliott")
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: GFP for selecting transgenic cells
Date: 11 May 1999 22:44:32 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 71
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <9905120537.AA05997@centaur>
NNTP-Posting-Host: net.bio.net

Hi all,

There has been recent interest in the use of GFP for identifying and
isolating transgenic plant cells for the recovery of transgenic plants
without conventional antibiotic or herbicide-based selection. We have
investigated this and there is a discussion on this topic in a Plant Cell
Reports paper, which is just coming out now. I am not sure whether
everybody will be able to access the web-based version and if not, I will
place a copy (when I can get it myself) on our ftp site. It would be
preferable and quicker to download the paper from there, then to ask for a
reprint request.

It is certainly possible to select transgenic cells and to recover GFP+
plants on the basis of visual scanning using a low power fluorescence
microscope. This is of course more labor intensive than antibiotic
selection, requiring time to scan the plates, although we have reduced this
time quite significantly by low power scanning for highly fluorescent
patches. The early screening for GFP+ events can significantly reduce your
tissue culture load at a very early stage. GFP may therefore be included in
many transformation experiments as a secondary marker. In fact, we found
GFP to be particularly useful in transformation experiments for many
reasons. 

A. R. Elliott, J. A. Campbell, B. Dugdale, R. I. S. Brettell, C. P. L.
Grof:
Green-fluorescent protein facilitates rapid in vivo detection of
genetically transformed plant cells
Plant Cell Reports 18 (1999) 9, 707-714
URL:
http://link.springer.de/link/service/journals/00299/bibs/9018009/90180707.ht
m

I will be happy to discuss the results or methodologies in the paper if you
wish to email me. 


Regards
Adrian

Dr. Adrian Elliott               Voice: +61-7-3214-2311
Research Scientist            Fax:   +61-7-3214-2288
CSIRO, Tropical Agriculture   
306 Carmody Rd, St Lucia, QLD, Australia. 4067
Email : adrian.elliott@tag.csiro.au





Original posting by Philippe:

Dear all,

How do you consider the following idea ?
A production of transgenic plants without selection (negative or positive)
procedure.

Thank you for your comments,

Philippe.
________________________________________

Philippe HERVE
UMR CNRS/UPS 5546 
Pôle de Biotechnologie Végétale
24 Chemin de Borde Rouge BP 17 Auzeville
31326 Castanet Tolosan - FRANCE

Tél. 33 (0)5 62 19 35 50
Fax. 33 (0)5 62 19 35 02
E-Mail herve@cict.fr

From owner-fluorescent@net.bio.net Mon May 10 23:00:00 1999
Path: biosci!UNIV-ROUEN.FR!Sebastien.Paris
From: Sebastien.Paris@UNIV-ROUEN.FR (Sebastien Paris)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Looking for GFP monoclonal antibody
Date: 11 May 1999 10:34:28 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 22
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <3.0.1.32.19990511193130.00910600@osiris>
NNTP-Posting-Host: net.bio.net

Dear GFPers,

I want to make cytohistological assays and I need a GFP monoclonal
antibody. Does anyone know where I can buy this ? Does anybody have
references where such assays were performed ?

Thanks in advance

seb


________________________________________________________

Sebastien PARIS
INSERM U295
Faculte de Medecine
22, Bd Gambetta
79183 Rouen cedex
Tel : +33 (0)2 35 14 82 80
Fax : +33 (0)2 35 14 82 37
e-mail : Sebastien.Paris@univ-rouen.fr
________________________________________________________

From owner-fluorescent@net.bio.net Mon May 10 23:00:00 1999
Path: biosci!internet!biosci!not-for-mail
From: biohelp (BIOSCI Administrator)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: BIOSCI/bionet miniFAQ & Fundraiser
Date: 11 May 1999 02:00:14 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 233
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <199905110900.CAA28931@net.bio.net>
NNTP-Posting-Host: net.bio.net

(LAST REVISION: 30-JUL-95)

This BIOSCI "miniFAQ" is designed to answer the questions that come up
the *most frequently*.  The main BIOSCI FAQ (Frequently Asked
Questions) is accessible on the World Wide Web at URL
http://www.bio.net/.

If you can not find an answer to your question in this or other
documentation, the BIOSCI technical support staff answers e-mail
queries sent to

		       biosci-help@net.bio.net

We can only answer questions about the use of the newsgroups and
mailing lists.  We unfortunately do not have the staff to do Internet
information searches or answer scientific questions.  Please post
those to the appropriate BIOSCI/bionet newsgroups.


	Contents:
	--------
	0) BIOSCI NEEDS YOUR SUPPORT!!

	1) Using the WWW to access the BIOSCI/bionet newsgroups.

	2) What to do about "spams," i.e., junk mail, ads, etc.

	3) Examples of subscribing and unsubscribing to the mailing lists.

	4) The BIOSCI user address and research interest directory.


0) BIOSCI NEEDS YOUR SUPPORT!!
------------------------------
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1) Using the WWW to access the BIOSCI/bionet newsgroups.
--------------------------------------------------------
As of 10 December 1995, all BIOSCI/bionet full newsgroups are
accessible through the World Wide Web (WWW) at URL http://www.bio.net.
One can read and reply publicly or privately to both recent postings
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-------------------------------------------------------
BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups),
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The same postings are distributed on all media (except for a small
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What should you do personally if you get junk mail?
---------------------------------------------------
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What can BIOSCI/bionet do to protect its newsgroups?
----------------------------------------------------
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We will moderate any of our newsgroups if the discussion leader tells
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Users in the Americas and Pacific Rim countries who use the BIOSCI
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To subscribe and unsubscribe to/from the BIOSCI lists, you need to
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-----------------------------------------------------------
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From owner-fluorescent@net.bio.net Tue May 11 23:00:00 1999
Path: biosci!WEIZMANN.WEIZMANN.AC.IL!cplachis
From: cplachis@WEIZMANN.WEIZMANN.AC.IL (Aurelie)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: preserving GFP
Date: 12 May 1999 05:26:12 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 8
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <373971FA.8175F76C@weizmann.weizmann.ac.il>
NNTP-Posting-Host: net.bio.net

Hi,
I would like to have some information about the conditions
(concentration, temperature, PH, etc.) in which GFP can be preserved for
long periods and not denature or aggregate.
Thanks, Aurelie.




From owner-fluorescent@net.bio.net Wed May 12 23:00:00 1999
Path: biosci!utkux.utcc.utk.edu!vonarnim
From: vonarnim@utkux.utcc.utk.edu
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: EGFPs in plants
Date: 13 May 1999 07:34:47 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 15
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <v01540b00b36090c95e41@[128.169.242.211]>
NNTP-Posting-Host: net.bio.net

Dear newsgroup: Has anyone of you had experience with expressing CLontech's
EGFPs in plant cells?  What were the results? Which cell type was used?
Thank you - Albrecht

Albrecht von Arnim

   Assistant Professor
   Dept. of Botany
   University of Tennessee
   Knoxville, TN 37996-1100
   vonarnim@utk.edu
   Phone: (423) 974-6206
   Fax:   (423) 974-0978



From owner-fluorescent@net.bio.net Wed May 12 23:00:00 1999
Path: biosci!CSHL.ORG!guest
From: guest@CSHL.ORG (CSHL Course Rental)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Photoconversion
Date: 13 May 1999 17:51:50 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 10
Sender: daemon@net.bio.net
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I am trying to photoconvert EGFP using DAB in brain tissue and so far
without success.  If anyone has any helpful advice I would be most
grateful.

Please reply to G.W.Knott@utas.edu.au


Thanks,
Graham


From owner-fluorescent@net.bio.net Wed May 12 23:00:00 1999
Path: biosci!CSHL.ORG!guest
From: guest@CSHL.ORG (CSHL Course Rental)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Photoconversion of GFP
Date: 13 May 1999 17:53:07 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 2
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <373B728C.D0006429@cshl.org>
NNTP-Posting-Host: net.bio.net

My email address is g.w.knott@utas.edu.au.


From owner-fluorescent@net.bio.net Sat May 15 23:00:00 1999
Path: biosci!INAME.COM!tmspro1
From: tmspro1@INAME.COM
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Does your business need to accept Credit Cards?
Date: 16 May 1999 18:53:59 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 49
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <B0000024032@ntserver.salford-col.ac.uk>
NNTP-Posting-Host: net.bio.net




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From owner-fluorescent@net.bio.net Sun May 16 23:00:00 1999
Path: biosci!iaf.cnrs-gif.fr!Andre.Klarsfeld
From: Andre.Klarsfeld@iaf.cnrs-gif.fr
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: (none)
Date: 17 May 1999 06:19:14 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 52
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <v03020900b365c2bda3caF@[157.136.26.86]>
NNTP-Posting-Host: net.bio.net

;, Fines Mouches:;, fglenne@aol.com, fluorpro@net.bio.net, rouyer,
 Francois.SCHACHTER@DeVinci.FR, f.revah@cerep.fr, gam@iap.fr,
 Genetique et Liberte:;, gel@altavista.net, Jacques.Stinnakre@cnrs-gif.fr,
 Perrier <jjp@imaginet.fr>, jlbesse@pasteur.fr (Jean-Louis Bessereau),
 jean-maurice.dura@emex.u-psud.fr,
 Jean-Paul Bouillot <bouillot@wat.nbcm.cnrs-gif.fr>, Keith@pasteur.fr,
 "Dr C.P. Kyriacou" <cpk@leicester.ac.uk>,
 MARC BALLIVET <Marc.Ballivet@biochem.unige.ch>,
 marie-aline.bloch@wanadoo.fr, pinto@igmors.u-psud.fr, pcossart@pasteur.fr,
 Patrice Muller <patrice.muller@hol.fr>, galison@husc.harvard.edu,
 pdaubas@pasteur.fr, psanson@pasteur.fr,
 Pierre-Henri.Gouyon@esv.u-psud.fr (Pierre Henri Gouyon),
 pilougoo@pasteur.fr, pouchele@vesinet.inserm.fr,
 rebource@tolbiac.inserm.fr, Salomon@infobiogen.fr, Spencer Brown <>,
 alonso@ibdm.univ-mrs.fr, Shaynoor.Dramsi@pasteur.fr,
 "G. et S. Klarsfeld" <gklarsfeld@minitel.fr>, spradling@mail1.ciwemb.edu,
 steph@genset.fr (Stephane Bejanin), dieudon@biologie.ens.fr,
 smichel@rz.uni-leipzig.de, burden@mcbi-34.med.nyu.edu (Steve Burden),
 vdavid@pasteur.fr
From: Andre Klarsfeld <Andre.Klarsfeld@iaf.cnrs-gif.fr>
Subject: PREVENTION ANTI-VIRALE

Bonjour,
Ce message est destiné à vous prévenir, au cas où vous recevriez un message
prétendument de ma part  (ou de n'importe qui d'autre) intitulé "IMPORTANT
MESSAGE FROM André Klarsfeld (ou FROM XXX)" :

N'OUVREZ PAS le fichier attaché Word qui l'accompagne. Mettez-le à la
corbeille.
Il contiendrait le virus Melissa, qui vous fera ensuite le même coup, à
savoir envoyer un message de votre part à 50 adresses email de votre
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Si vous n'ouvrez pas le fichier attaché Word, le virus, qui est une Macro
Word, ne peut pas infecter votre ordinateur.

Désolé pour ce tracas,
André


_______________________________________________
Andre Klarsfeld
"Genetique moleculaire des rythmes circadiens"
CNRS Inst. A. Fessard
91198 GIF-SUR-YVETTE cedex
tel : 33-(0)1 69 82 43 04
fax : 33-(0)1 69 07 05 38
email : Andre.Klarsfeld@iaf.cnrs-gif.fr

http://www.cnrs-gif.fr/iaf/iaf3/equipe3.html



From owner-fluorescent@net.bio.net Mon May 17 23:00:00 1999
Path: biosci!tag.csiro.au!adrian.elliott
From: adrian.elliott@tag.csiro.au ("Adrian Elliott")
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: HIS-tagged GFP supplier ?
Date: 18 May 1999 01:00:29 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 15
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <9905180754.AA04147@centaur>
NNTP-Posting-Host: net.bio.net

Hi,

Clontech does not seem to be supplying the HIS-tagged GFP any longer. Can
somebody supply a plasmid or point me towards another supplier ?

Regards
Adrian

Dr. Adrian Elliott               Voice: +61-7-3214-2311
Research Scientist            Fax:   +61-7-3214-2288
CSIRO, Tropical Agriculture   
306 Carmody Rd, St Lucia, QLD, Australia. 4067
Email : adrian.elliott@tag.csiro.au



From owner-fluorescent@net.bio.net Mon May 17 23:00:00 1999
Path: biosci!hassle.se.astra.com!vijay.chhajlani
From: vijay.chhajlani@hassle.se.astra.com
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: receptor-GFP fusion
Date: 18 May 1999 01:01:49 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 16
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <3DB0284E2B7AD21189A400805F19DB288C4A26@se-hsl-mail010.hassle.se.astra.com>
NNTP-Posting-Host: net.bio.net

Hi,
I would like to prepare a GPCR and GFP fusion protein. I would like to fuse
GFP at N and C terminal of receptor in two different constructs. Can any one
help me with idead for which of the GFP variants are best for studying
receptor sequestration and internalisation, preferably in real time. Are
there any kits available for making such constructs.
Regards
Vijay Chhajlani,
HB4
Cell Biology & Biochemistry
Astra Hässle AB
431 83 Mölndal
Sweden
Tel: 46 31 776 2097
Fax: 46 31 776 3761
Vijay.chhajlani@hassle.se.astra.com

From owner-fluorescent@net.bio.net Mon May 17 23:00:00 1999
Path: biosci!NS1.SUBMISSIONSERVICE.COM!sales
From: sales@NS1.SUBMISSIONSERVICE.COM
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: ADV: your web site
Date: 18 May 1999 11:59:58 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 75
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <199905181859.LAA26732@net.bio.net>
NNTP-Posting-Host: net.bio.net

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From owner-fluorescent@net.bio.net Wed May 19 23:00:00 1999
Path: biosci!news.stanford.edu!newsfeed.stanford.edu!logbridge.uoregon.edu!dispose.news.demon.net!demon!newsfeed.tli.de!newscore.gigabell.net!newscore.ipf.de!news-fra1.dfn.de!news-lei1.dfn.de!news.uni-leipzig.de!news.uni-halle.de!not-for-mail
From: Marcus Wernitz <viacx@mlucom6.urz.uni-halle.de>
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Re: HIS-tagged GFP supplier ?
Date: Thu, 20 May 1999 15:29:59 +0200
Organization: University Halle, Germany
Message-ID: <37440E57.4069ECD8@mlucom6.urz.uni-halle.de>
References: <9905180754.AA04147@centaur>
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Hi Adrian,

If you look for the plasmid with GFP and 6xHis, Igot a very useful adress for
you:

http://gillnet.lab.nig.ac.jp/~cvector/NIG_cvector/aboute.html

They ship you lots of different plasmids for free, as long as you tell them
what you need the plasmid for.
Just try it there.

Good luck,
Marcus


Adrian Elliott schrieb:

> Hi,
>
> Clontech does not seem to be supplying the HIS-tagged GFP any longer. Can
> somebody supply a plasmid or point me towards another supplier ?
>
> Regards
> Adrian
>
> Dr. Adrian Elliott               Voice: +61-7-3214-2311
> Research Scientist            Fax:   +61-7-3214-2288
> CSIRO, Tropical Agriculture
> 306 Carmody Rd, St Lucia, QLD, Australia. 4067
> Email : adrian.elliott@tag.csiro.au


From owner-fluorescent@net.bio.net Thu May 20 23:00:00 1999
Path: biosci!UMDNJ.EDU!liuhy
From: liuhy@UMDNJ.EDU
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: EGFP under UV
Date: 21 May 1999 14:48:21 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 11
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NNTP-Posting-Host: net.bio.net

i am trying to compare wt GFP and EGFP fusion proteins expression in
yeast. i "thought" that EGFP only has 1 excitation peak which ranges in
490 nm, so that when i used UV filter, i should NOT see any signal. but
i did see green color with EGFP fusion protein when i used UV filter,
although it is less light than the wtGFP. could anyon ehelp me to figure
out this??? or i got the wrong impression? what is the difference of
wtGFP v.s. EGFP???

thank you very much

hsing-yin

From owner-fluorescent@net.bio.net Sat May 22 23:00:00 1999
Path: biosci!COLUMBIA.EDU!tw118
From: tw118@COLUMBIA.EDU (tomoko watanabe)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Is magenta out there?
Date: 23 May 1999 20:46:20 -0700
Organization: columbia university
Lines: 9
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NNTP-Posting-Host: net.bio.net

Hi!

I heard that a few groups have began to use magenta fluorescent
protein.  Is it true that magenta protein exists?  If so, is it
commercially available?  What'sthe emmission spectra, etc?  Any
information would be greatly appreciated...Thanks!

tomoko watababe


From owner-fluorescent@net.bio.net Sun May 23 23:00:00 1999
Path: biosci!news.stanford.edu!newsfeed.stanford.edu!logbridge.uoregon.edu!feeder.qis.net.MISMATCH!feeder.qis.net!nntp.abs.net!newshub2.home.com!news.home.com!news.rdc1.az.home.com.POSTED!not-for-mail
From: "george sibbald" <geos@goldrush.com>
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: Poster / Images:  Protein folding force characterization via AFM
Lines: 15
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Date: Mon, 24 May 1999 23:03:37 GMT
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Poster / Images:  Protein folding force characterization via AFM

Biological Measurements and Forces in
MAC Mode™ AFM
Stuart Lindsay, Wenhai Han, and Yanzhang Liu; ASU

Biological force measurements and the AFM
Protein unfolding may play a vital role in protein function (Soteriou,
Clarke et al. 1993). Last year, Rief et al. (Rief, Gautel et al. 1997)
demonstrated that the AFM could be used to follow the unfolding of a single
protein molecule trapped between the AFM probe and a gold surface

http://www.molec.com/newsletters/spring98/npage1.htm



From owner-fluorescent@net.bio.net Mon May 24 23:00:00 1999
Path: biosci!news.stanford.edu!newsfeed.stanford.edu!remarQ73!supernews.com!remarQ.com!newsfeed.berkeley.edu!usenet.INS.CWRU.Edu!not-for-mail
From: Scott Vande Pol <sbv@po.cwru.edu>
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: GFP-neo fusions active?
Date: Tue, 25 May 1999 12:37:33 -0400
Organization: CWRU
Lines: 5
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Dear Colleagues

Has anyone made a GFP-neo fusion that is active for both fluorescence
and neo selection?


From owner-fluorescent@net.bio.net Mon May 24 23:00:00 1999
Path: biosci!news.stanford.edu!newsfeed.stanford.edu!newsfeed.berkeley.edu!usenet.INS.CWRU.Edu!not-for-mail
From: Scott Vande Pol <sbv@po.cwru.edu>
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: GFP-neo fusions active?
Date: Tue, 25 May 1999 12:37:05 -0400
Organization: CWRU
Lines: 5
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Dear Colleagues

Has anyone made a GFP-neo fusion that is active for both fluorescence
and neo selection?


From owner-fluorescent@net.bio.net Mon May 24 23:00:00 1999
Path: biosci!news.stanford.edu!newsfeed.stanford.edu!news.ems.psu.edu!news.cis.ohio-state.edu!nntp.service.ohio-state.edu!usenet.INS.CWRU.Edu!not-for-mail
From: Scott Vande Pol <sbv@po.cwru.edu>
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: HyTk or Tkneo selections markers
Date: Tue, 25 May 1999 12:27:32 -0400
Organization: CWRU
Lines: 9
Message-ID: <374ACF74.A14237C@po.cwru.edu>
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Dear Colleagues
I am searching for the positive / negative selection  markers HygTK
(hygromycin/Thymidine kinase fusion) and the TKneo (thymidine kinase
fused to neomycin resistance gene).  If anyone has one or both oth these
or knows a ready source please email : sbv@po.cwru.edu.


Thanks for any help!


From owner-fluorescent@net.bio.net Tue May 25 19:28:00 1999
Path: biosci!MINDSPRING.COM!grizzlyan
From: grizzlyan@MINDSPRING.COM (Michael Sherrell)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: MS, seqs, synths, NMRs for sale
Date: 25 May 1999 20:28:46 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 41
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Distribution: world
Message-ID: <01BEA6EC.C9F19CE0@user-33qsohi.dialup.mindspring.com>
NNTP-Posting-Host: net.bio.net

Mass spectrometers, sequencers, synthesizers, NMRs for sale:
LC/Mass spectrometers:
     Sciex API III+ LC/MS/MS $ 69,000 ES, APCI, under PE service contract
     HP 1100 benchtop LC/MSD, APCI & API-electrospray, autosampler, DAD detector, < 1 year old, $130,000
     Finnigan Navigator benchtop LC/MS: $ 75,000; 18 mos. old; factory refurb, install & 90-day warr. included
    Finnigan TSQ 7000 LC+GC/MS/MS: $ 97,500; current software; API-1 source; under Finnigan service contract
     Sciex 150 benchtop LC/MS: $ 98,000; 2 yrs old; includes Gilson 215 liquid handler, ELSD detector & HP 1100 HPLC.
      Finnigan SSQ 7000 LC/MS: $130,000; API-2 source, electrospray + APCI, Excaliber software, factory refurb, 90-day warranty, install included
     Micromass II: $50,000; ES, APCI, 3 yrs. old, includes HPLC, NOT Z-spray
     HP 5989B LC+GC/MS: $ 35,000; Extended mass range (2000 amu), hex ion guide; HPLC & warranty avail.
     Finnigan MAT 90: $30,000; Hi-resolution magnetic sector
     Fisons VG 2000: <$100,000
MALDI-TOFs:
     Hewlett-Packard G2025A $ 50,000
Peptide and oligo synthesizers and sequencers:
     ABI 394: $12,500 (Valve blocks rebuilt; warranteed)
     ABI 390Z: $4,000 (50-100uM yields)
     ABI 431: $12,500 (Rebuilt, warranteed)
     ABI 433: $19,000 (ABI upgrade)
     PerSeptive 9050+: $6,000 (As is/was working when decommissioned; add $3,500 for rebuild/warranty)
     ABI 373 stretch: $9,000 (Big dye upgrade; still under warranty)
     ABI 373 stretch: $7,000 (4-filter)
     ABI 377: $97,500 (XL; 96 lanes; transferrable ABI service contract)
     ABI 377: $60,000 (48-lane; under ABI contract)
     ABI Procise 492: $45,000 obo (ABI-certified)
NMRs:
     Bruker DMX 600: $495,000; AVANCE; 3-axis gradients
     Varian Unity 500+: call for price; broadband
     Bruker AM360: $85,000; broadband; widebore; install included
     Varian Gemini 300: call for price; widebore; broadband; remote sampling arm; ~1992
     Varian Gemini 300: $69,000; proton/carbon probe; freight, install, 90-day warr. included
     Varian Unity 500: call for price; broadband
Also available:
      Hitachi 570 scanning electron microscope, Kevex detector, running now, $35,000.
      BD FACSVantage and assorted FACScans
Various other seqencers, synthesizers etc. are available; please inquire or check the website.

Michael Sherrell
Grizzly Analytical
707 887 2919/fax 707 887 9834
www.grizzlyanalytical.com

From owner-fluorescent@net.bio.net Wed May 26 01:41:00 1999
Path: biosci!AOL.COM!9822
From: 9822@AOL.COM
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: ARE YOU BEING CHEATED ON??
Date: 26 May 1999 02:41:58 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 33
Sender: daemon@net.bio.net
Distribution: world
Message-ID: <199905260835.KAA09088@linux2.rz.fh-hannover.de>
NNTP-Posting-Host: net.bio.net



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From owner-fluorescent@net.bio.net Wed May 26 12:41:00 1999
Path: biosci!UMDNJ.EDU!liuhy
From: liuhy@UMDNJ.EDU
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: fixation of GFP-expressing yeast
Date: 26 May 1999 13:41:10 -0700
Organization: BIOSCI International Newsgroups for Molecular Biology
Lines: 12
Sender: daemon@net.bio.net
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Message-ID: <374C5C45.1185@rwja.umdnj.edu>
Reply-To: liuhy@UMDNJ.EDU
NNTP-Posting-Host: net.bio.net

Thanks people in this grop gave me so much valuable suggestion for the
last question i asked.  Now i am facing another problem, I tried to fix
my yeast cells with 4% of paraformaldehyde like Clontech suggested in
the handbook, sometimes i got good results, but lots of times i just got
reduced GFP signal after fixed for 15 or 30min in this solution.  I
wonder if anybody out there tried to fix yeast cells expressing GFP
fusion protein???( i have lots of samples and our lab don't have the
microscope, i don't think using fresh cells is an option for me)

thank you very for your help in advance.

hsing-yin

From owner-fluorescent@net.bio.net Wed May 26 23:01:00 1999
Path: biosci!rutgers!rockyd.rockefeller.edu!newsfeed.nyu.edu!news.idt.net!newsfeed.cwix.com!202.54.1.25!news.vsnl.net.in!not-for-mail
From: "Anisha" <anisha_99@usa.net>
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: YOUNG BEAUTIFUL MODEL AVAILABLE
Date: Thu, 27 May 1999 12:35:41 +0530
Organization: VSNL
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------=_NextPart_000_003B_01BEA83D.6E7E6080--


From owner-fluorescent@net.bio.net Thu May 27 13:07:00 1999
Path: biosci!news.stanford.edu!newsfeed.stanford.edu!logbridge.uoregon.edu!newsrelay.iastate.edu!news.iastate.edu!pourel
From: pourel@iastate.edu (I. Pour-El)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: GFP toxicity
Date: 27 May 1999 20:57:32 GMT
Organization: Iowa State University, Ames, Iowa USA
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NNTP-Posting-Host: isua2.iastate.edu

Anyone have any information on GFP or GFP fragment overproduction toxicity
in E coli?

I. Pour-El
Iowa State University

-- 
I. Pour-El						pourel@iastate.edu
Iowa State University					
Ames, IA 50011

From owner-fluorescent@net.bio.net Sat May 29 00:32:00 1999
Path: biosci!COMPUSERVE.COM!642112.702
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e@sonic.net, fluorite@msn.com, fluoroborate@rootwisem.int, fluorpro@net.bio.net, fluoxetine@pharmacy.com, flup@flop.com, flup@terra.sirius.com, flup@worldonline.nl, flupis1@hotmail.com
From: <642112.702@compuserve.com>
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From owner-fluorescent@net.bio.net Sun May 30 15:59:00 1999
Newsgroups: bionet.molbio.proteins.fluorescent
Path: biosci!agate!newsfeed.berkeley.edu!news.maxwell.syr.edu!nntp.news.xara.net!xara.net!server6.netnews.ja.net!leeds.ac.uk!news
From: bmbjmm@bmb.leeds.ac.uk (Jeremy Murray)
Subject: Re: Is magenta out there?
X-Accept-Language: en
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there is a calcium binding,
blue fluorescent protein
check out Chemistry & Biology, (1996), 3(5)pp337-47
hth, jez

tomoko watanabe wrote:

> Hi!
>
> I heard that a few groups have began to use magenta fluorescent
> protein.  Is it true that magenta protein exists?  If so, is it
> commercially available?  What'sthe emmission spectra, etc?  Any
> information would be greatly appreciated...Thanks!
>
> tomoko watababe


From owner-fluorescent@net.bio.net Mon May 31 06:42:00 1999
Path: biosci!newshost.lanl.gov!logbridge.uoregon.edu!dispose.news.demon.net!demon!colt.net!news0.de.colt.net!bignews.mediaways.net!fu-berlin.de!server1.netnews.ja.net!server2.netnews.ja.net!hgmp.mrc.ac.uk!biosci
From: Wolfgang.Schechinger@med.uni-tuebingen.de ("Wolfgang Schechinger")
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: making movies out of multi-image tif files
Date: 31 May 1999 15:36:46 +0100
Organization: MRC Human Genome Mapping Project Resource Centre
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Hi all, 

I have some thick image stacks (2048*2048 pixel, 50 images) I would 
like to convert into *.avi files or maybe better into jpeg encoded 
(or otherwise compressed) movie files. Is there any software 
(freeware or shareware preferred but not necessarily) out there that 
will aid me in this purpose? I have access to NT and linux / UNIX 
environments. 

All input is warmly welcome.

Wolfgang
-----
usual disclaimers apply * This message is RNAse free - please don't touch!
-----                              
Wolfgang Schechinger   
Pathobiochemistry Dept.      
University of Tuebingen, Germany
email: wgschech@med.uni-tuebingen.de * wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
-------
*unsolicited mail is *NOT* appreciated
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From owner-fluorescent@net.bio.net Mon May 31 20:38:00 1999
Path: biosci!DMSNA.DMS.UNINA.IT!mey81
From: mey81@DMSNA.DMS.UNINA.IT
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: ADV: Photo Mousepads......These are cool!
Date: 31 May 1999 21:38:22 -0700
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To be removed from our mailing list please call 800-242-0363 ext. 2759
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From owner-fluorescent@net.bio.net Mon May 31 22:43:00 1999
From: t.humphrey@uq.edu.au (Tania Humphrey)
Newsgroups: bionet.molbio.proteins.fluorescent
Subject: GFP FRET in plants
Date: 1 Jun 1999 06:37:45 GMT
Organization: University of Queensland
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Hi

Has anyone out there done Fluorescent Resonance Energy Transfer (FRET) with 
GFP variants in plants especially Arabidopsis?  Any references, names or other 
information would be a great help.

Thankyou

Tania Humphrey

Department of Botany
University of Queensland
Australia