From owner-fluorescent@net.bio.net Mon Dec 02 22:00:00 1996 Path: biosci!ihnp4.ucsd.edu!swrinde!cs.utexas.edu!newshost.convex.com!newsgate.duke.edu!news-server.ncren.net!concert!ussun2n.glaxo.com!usenet From: Sam Witherspoon Newsgroups: bionet.molbio.proteins.fluorescent Subject: Relative fluorescence of GFP / molar fluorescence yield Date: Tue, 03 Dec 1996 17:00:37 -0800 Organization: Glaxo Wellcome Inc. Lines: 16 Message-ID: <32A4CD35.2350@usav01.glaxo.com> NNTP-Posting-Host: us4l76.glaxo.com Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.02 (Win16; U) CC: sw11527@glaxo.com frank parlati wrote: > > Does anyone know how many GFP molecules per unit volume (in particular the > S65T and Y66H/Y145F mutants) are required to observe a signal by > microscopy. Along the same lines, does anyone have any information on molar fluorescence yield for GFP of GFP S65T? How does it compare to fluorescein on a per molecule basis? Thanks ------------------------------------------------------------------------Sam Witherspoon e-mail: sw11527@glaxo.com Flow Cytometry and Cell Sorting Resource Tel: 919-483-3078 Dept. of Receptor Biochemistry FAX: 919-483-3777 Glaxo Wellcome R&D 5 Moore Dr. RTP, NC, 27709 USA From owner-fluorescent@net.bio.net Mon Dec 02 22:00:00 1996 Path: biosci!MRI8.MRI.MONTANA.EDU!jay From: jay@MRI8.MRI.MONTANA.EDU (jay brewster) Newsgroups: bionet.molbio.proteins.fluorescent Subject: autofluorescence Date: 3 Dec 1996 14:44:10 -0800 Organization: McLaughlin Institute for Biomedical Research Lines: 7 Sender: daemon@net.bio.net Distribution: world Message-ID: <32A4AEA1.1682@po.mri.montana.edu> NNTP-Posting-Host: net.bio.net Newsnet users; I have a reporter mouse line expressing EGFP to characterize a promoter. I am getting good signal in subsets of cells in several tissues. I am using a HiQ FITC cube and am getting quite a bit of autofluorescence in some tissue. What is the best way to deal with this? (another filter?, an addition step to eliminate the autofluorescent?). Also what is the best way to fix and mount tissue for GFP, I seem to be getting photobleaching in Fluorosave. Thanks! jay@po.mri.montana.edu From owner-fluorescent@net.bio.net Mon Dec 02 22:00:00 1996 Path: biosci!USAV01.GLAXO.COM!nfc20355%usa.decnet From: nfc20355%usa.decnet@USAV01.GLAXO.COM (Neal Cariello) Newsgroups: bionet.molbio.proteins.fluorescent Subject: GFP dimerization Date: 3 Dec 1996 09:53:01 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 14 Sender: daemon@net.bio.net Distribution: world Message-ID: <32A49284.50A4@usav01.glaxo.com> NNTP-Posting-Host: net.bio.net Hello, I know that GFP can dimerize at very high concentrations, like those used in crystallization. Does anyone know if GFP dimerizes at physiolgical concentrations? My real question is: If dimerization occurs would the dimer fluoresce if one molecule was functional and the other was not? Any thoughts are welcome. Regards, Neal Cariello From owner-fluorescent@net.bio.net Tue Dec 03 22:00:00 1996 Path: biosci!agate!howland.erols.net!cam-news-hub1.bbnplanet.com!news.bbnplanet.com!su-news-hub1.bbnplanet.com!arclight.uoregon.edu!news.bc.net!unixg.ubc.ca!lpss From: lpss@unixg.ubc.ca (Alex Chang) Newsgroups: bionet.molbio.proteins.fluorescent Subject: Green fluorescent protein vector Date: 4 Dec 1996 01:41:31 GMT Organization: University of British Columbia, Vancouver, B.C., Canada Lines: 11 Message-ID: <582ksb$f01@nntp.ucs.ubc.ca> NNTP-Posting-Host: netinfo.ubc.ca X-Newsreader: TIN [version 1.2 PL2] Dear Netters, I have a quick question: Is that neccessary to put a linker sequence in between the protein intended to express and the GFP (in a CMV promoter vector, GFP at the C'terminus)? -- Alex Chang Pathology University of British Columbia achang@hivnet.ubc.ca From owner-fluorescent@net.bio.net Tue Dec 03 22:00:00 1996 Path: biosci!ihnp4.ucsd.edu!swrinde!howland.erols.net!news-peer.gsl.net!news.gsl.net!news-dc.gsl.net!news.gsl.net!news.NetVision.net.il!news From: Martin Sanders Newsgroups: bionet.molbio.proteins.7tms_r,bionet.molbio.proteins.fluorescent,bionet.molecules.peptides Subject: Optimize protein HPLC by monitoring conductivity and pH Date: 4 Dec 1996 18:29:23 GMT Organization: NetVision LTD. Lines: 14 Message-ID: <584fu3$o9v@news.NetVision.net.il> NNTP-Posting-Host: ts002p12.pop2a.netvision.net.il Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.2N (Windows; I; 16bit) Xref: biosci bionet.molbio.proteins.7tms_r:945 bionet.molbio.proteins.fluorescent:854 bionet.molecules.peptides:547 You can improve the separation of proteins and other biopolymers in HPLC systems for ion exchange, hydroxylapatite, and hydrophobic interaction by monitoring the conductivity of the eluent stream using an inexpensive flow through conductivity monitor. Proteins normally elute at the same conductivity for each run and monitoring conductivity of each run can therefore provide evidence that chromatographic conditions have been reproduced while giving an accurate picture of the salt gradient for each run. Can also be used to monitor HPLC column regeneration and monitor desalting of proteins by gel filtration. Monitoring of eluent pH is also discussed. For further details contact Lazar Research Labs. Inc. by emailing service@lazarlab.com or faxing 1-213-931-1434 or viewing the Lazar web site at http://www.lazarlab.com From owner-fluorescent@net.bio.net Tue Dec 03 22:00:00 1996 Path: biosci!U.WASHINGTON.EDU!moser From: moser@U.WASHINGTON.EDU ("'Mike' Michael J. Moser") Newsgroups: bionet.molbio.proteins.fluorescent Subject: Re: autofluorescence Date: 4 Dec 1996 08:53:04 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 25 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <32A4AEA1.1682@po.mri.montana.edu> NNTP-Posting-Host: net.bio.net Jay, I have noticed autofluorescence in live or fixed HT1080 fibroblast cells that I use. This fluorescence can be distinguished from that of GFP because it is still visible with the rhodamine cube. It must have a broader spectrum of emmission than GFP. It's still a pain, but at least you can differentiate GFP from bkg fluorescence. MIKE Mike Moser Tel: 206-616-7391 UW Department of Pathology FAX: 206-543-3644 Box 357470 moser@u.washington.edu Seattle, WA 98195 http://weber.u.washington.edu/~moser On 3 Dec 1996, jay brewster wrote: > Newsnet users; > I have a reporter mouse line expressing EGFP to characterize a promoter. I am getting > good signal in subsets of cells in several tissues. I am using a HiQ FITC cube and am > getting quite a bit of autofluorescence in some tissue. What is the best way to deal > with this? (another filter?, an addition step to eliminate the autofluorescent?). > Also what is the best way to fix and mount tissue for GFP, I seem to be getting > photobleaching in Fluorosave. Thanks! jay@po.mri.montana.edu > > From owner-fluorescent@net.bio.net Wed Dec 04 22:00:00 1996 Path: biosci!ZENTRUM.PHYS.CHEMIE.TU-MUENCHEN.DE!harald From: harald@ZENTRUM.PHYS.CHEMIE.TU-MUENCHEN.DE (Harald Lossau) Newsgroups: bionet.molbio.proteins.fluorescent Subject: RE: GFP Dimerization Date: 5 Dec 1996 09:05:57 -0800 Organization: TU-Muenchen Lines: 11 Sender: daemon@net.bio.net Distribution: world Message-ID: <32A77FD9.17C7@zentrum.phys.chemie.tu-muenchen.de> NNTP-Posting-Host: net.bio.net Neal, it has been suggested that GFP is dimeric in vivo (M.W.Cutler and W.Ward, Photochem. Photobiol. 57, 63S (1993). However, dimerization is not necessary for GFP to fluoresce. Time resolved fluorescence is not influenced by cryoprotectors which supress dimerization (H. Lossau et al., Chemical Physics 2653 (Dec. 1996)). Harald Lossau From owner-fluorescent@net.bio.net Sun Dec 08 22:00:00 1996 Path: biosci!agate!howland.erols.net!www.nntp.primenet.com!nntp.primenet.com!news.mindspring.com!mindspring!uunet!in2.uu.net!192.109.159.3!news.gtn.com!inn.aball.de!on-line.leine.de!yamato.leine.de From: mapstool@B-52.zer Newsgroups: bionet.molbio.proteins.fluorescent Subject: Autoinit-Mail Date: 8 Dec 96 01:51:44 +0100 Message-ID: <239DD22023MM001@mapstool.chatline.zer> X-Gateway: ZCONNECT UH online.leine.de [UUCPfZ V5.81 U055] Lines: 2 ! From owner-fluorescent@net.bio.net Sun Dec 08 22:00:00 1996 Path: biosci!bio.tokushima-u.ac.jp!yoshioka From: yoshioka@bio.tokushima-u.ac.jp (hidefumi yoshioka) Newsgroups: bionet.molbio.proteins.fluorescent Subject: pEGFP vector can be express in Drosophila embryo Date: 9 Dec 1996 18:47:32 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 16 Sender: daemon@net.bio.net Distribution: world Message-ID: <9612100242.AA27628@bio.bio.tokushima-u.ac.jp> NNTP-Posting-Host: net.bio.net >Did anybody succeed in expression of GFP fusion with a pEGFP vector >(clonetech) in Drosophila embryo ? I would like to know what GFP vector is suitable in Drosophila embryo. > > !!!!!!!!-------------------------------------------------------------------- ------------------!!!!!!!!! 吉岡　秀文 Hidefumi Yoshioka 徳島大学工学部 The University of Tokushima 生物工学科 　　　　 Faculty of Engineering Biological Science and Technology TEL +81 886-56-7529 FAX +81 886-55-3169 E-mail yoshioka@bio.tokushima-u.ac.jp From owner-fluorescent@net.bio.net Mon Dec 09 22:00:00 1996 Path: biosci!Bayer.com!Yinghe.Hu.B From: Yinghe.Hu.B@Bayer.com (Yinghe Hu) Newsgroups: bionet.molbio.proteins.fluorescent Subject: ligand-receptor interaction Date: 10 Dec 1996 06:59:22 -0800 Organization: Bayer Corporation Lines: 4 Sender: daemon@net.bio.net Distribution: world Message-ID: <32ADA4DF.7CAC@Bayer.com> NNTP-Posting-Host: net.bio.net Does a peptide ligand-GFP fusion protein interact with the peptide receptor? If so, will this interaction affect the level of GFP fluorescence? Any suggestion will be very much appreciated. From owner-fluorescent@net.bio.net Mon Dec 09 22:00:00 1996 Path: biosci!WELCHLINK.WELCH.JHU.EDU!kgupta From: kgupta@WELCHLINK.WELCH.JHU.EDU (KALPANA GUPTA) Newsgroups: bionet.molbio.proteins.fluorescent Subject: Charity (fwd) Date: 10 Dec 1996 12:42:07 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 31 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Hello all, This seems like one easy way to spread the spirit of the season. Wishing you all a Happy December. -Kalpana ---------- Forwarded message ---------- >From Wellesley's Public Bulletin: The Houghton-Mifflin publishing Co. is giving books to children's hospitals; how many books they give depends on how many emails they receive from people around the world. For every 25 emails they receive, they give one book--it seems like a great way to help a good cause. All you have to do is email share@hmco.com. Hope you can spare the seconds...and let your friends know. So far they only have 3, 400 messages...last year they reached 23,000. Again, the address is share@hmco.com (Share The Spirit Campaign) Info is also available at: http://www.polarexpress.com Happy holidays! From owner-fluorescent@net.bio.net Tue Dec 10 22:00:00 1996 Path: biosci!internet!biosci!not-for-mail From: biohelp (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins.fluorescent Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 11 Dec 1996 02:00:43 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 239 Sender: daemon@net.bio.net Distribution: world Message-ID: <199612111000.CAA22901@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!! ------------------------------ BIOSCI's government funding has been expended, and we are now operating solely from advertising revenue that we have raised from our Web site at http://www.bio.net/. We need just a few minutes of your time to help us serve you. You can do two important things which will take very little time for you individually and will immensely help us continue to help you. First, please use our WWW system at http://www.bio.net/ to access the archives. You can post or reply to messages via your Web browser as described in item #1 below. Your usage helps attract sponsors. If you contact any of our sponsors, please be sure to thank them for supporting BIOSCI. It is critical for them to get this feedback if they are to continue their sponsorship for the long term. Second, if you work for a company or organization that provides products or services of interest to the biology community, please pass this message on to your marketing or marketing communications department or other appropriate group. Please ask them to help support BIOSCI by sponsoring our Web site and explain the uses and benefits of the system to the biology community. If they are interested, they can then contact us for further information at our tech support address, biosci-help@net.bio.net. 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 and archived messages through one's Web browser if it is configured properly to send e-mail. Each newsgroup is equipped with its own WAIS index. The main BIOSCI home page also has access to the BIO-JOURNALS Table of Contents database WAIS index and the BIOSCI user address database described in another item further below. 2) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups), mailing lists, and a hypermail archive at URL http://www.bio.net/. The same postings are distributed on all media (except for a small number of mailing-list-only groups at net.bio.net). Unfortunately it is becoming a despicable practice on the Internet (by a few people out to make a fast buck) to do automated mass postings to thousands of newsgroups and mailing lists. These attempts to grab free advertising are refered to as "spams" in the usual, somewhat boneheaded, net terminology. USENET is more susceptible to this practice, and many spams originate on the USENET groups and then are passed on to the mailing lists. However, spammers also get lists of mailing addresses and hit these too, so neither medium is immune. What should you do personally if you get junk mail? --------------------------------------------------- Just delete it and move on without reading it further. Filing a protest is becoming increasingly useless because spammers are often disguising the addresses where the messages are sent from. Unless you really understand Internet mail systems, your attempt at protest by sending replies to the message will often end up being sent to the address of an innocent person that the spammer is victimizing. What can BIOSCI/bionet do to protect its newsgroups? ---------------------------------------------------- The only solution currently available is to moderate the newsgroup. If this newsgroup is already moderated, then you are in good shape. Moderation protects the USENET distribution from about 95% of the spams that are being sent to date and protects the mailing lists completely. Moderation means, however, that someone has to take the time to review each message before it goes out. We have set up software here that simply allows the moderator to forward to an address at net.bio.net messages that (s)he wishes to have distributed. This takes no more time than that needed to read the message and pass it on, say about 1 min. per message. Most newsgroups currently have a discussion leader who is responsible for their newsgroup. The discussions leaders and their e-mail addresses are listed in the BIOSCI Information Sheet which is available on the Web at http://www.bio.net/. If a newsgroup is being hit with too many junk postings, please contact the discussion leader for that group and see if there is interest in moderating the group. Please do not assume that by simply posting a complaint to the newsgroup itself, anyone on the BIOSCI staff will act on your complaint. With close to 100 newsgroups to run, the BIOSCI staff has to rely on the discussion leaders of each newsgroup to report problems directly to us at biosci-help@net.bio.net. We will moderate any of our newsgroups if the discussion leader tells us that the readership of the group wishes to do so and if a moderator is willing to do the work. For most BIOSCI/bionet groups, this entails only a few minutes of work each day. Moderating a newsgroup will resolve probably 95% of the junk postings on the USENET distribution. Unfortunately there are easy ways for determined spammers to override the moderation mechanism on USENET, but we can protect our e-mail subscribers from unwanted postings if the newsgroup is moderated. You can also access our newsgroups over the WWW at URL http://www.bio.net. While this Web interface will not stop spammers from trying to post to the groups, this will give you yet another way, besides using USENET news, to keep the junk out of your personal mail files. For those of you with local USENET news systems, the Web interface will also give you faster access to new newsgroups and recent postings. 3) Examples of subscribing and unsubscribing to the mailing lists. ------------------------------------------------------------------ PLEASE NOTE: The BIOSCI management does NOT act on subscription/unsubscription requests that are posted improperly to the newsgroups and mailing lists. People who do this only bother everyone on the lists to no avail. Please be sure to follow the proper procedures below. Gory details are in the BIOSCI Information sheets on the Web at http://www.bio.net. Below we give an example utilizing the METHODS-AND-REAGENTS list at both of our two BIOSCI sites: Users in the Americas and Pacific Rim countries who use the BIOSCI ------------------------------------------------------------------ node at computer net.bio.net: ---------------------------- A) Determine the "listname" which is the <=8 character mail address ^^^^^^^^^^^^^ for the group. These can be found in the BIOSCI Info. Sheet. For the METHODS-AND-REAGENTS group the mailing address is methods@net.bio.net. The listname is the portion of the address to the left of the @ sign, i.e., "methods". The listname is used with the "subscribe" and "unsubscribe" commands illustrated below. B) Mail all commands in the body of a mail message addressed to biosci-server@net.bio.net. Do NOT send commands to the newsgroup posting addresses! Leave the Subject: line blank, any text on it will be ignored. C) In the body of your message put one or more of the following commands with an "end" command on the last line, e.g., subscribe methods unsubscribe methods end Do NOT put your e-mail address or other text on these lines. The server only allows you to cancel your subscription if the address on your mail header matches the address on our mailing list. Please ask for help at biosci-help@net.bio.net if your address has changed, e.g., if you know you are on the list but the server tells you that you are not a member. Users in Europe, Africa, and Central Asia who use the BIOSCI node at -------------------------------------------------------------------- computer daresbury.ac.uk (also known as dl.ac.uk): ------------------------------------------------- To subscribe and unsubscribe to/from the BIOSCI lists, you need to specify the full USENET newsgroup name with "bionet-news." prepended. The USENET newsgroup names are listed in the BIOSCI Information sheet on the Web at http://www.bio.net/. For the METHODS-AND-REAGENTS list the USENET newsgroup name is bionet.molbio.methds-reagnts, thus the appropriate commands are sub bionet-news.bionet.molbio.methds-reagnts unsub bionet-news.bionet.molbio.methds-reagnts These commands are included in a message addressed to mxt@dl.ac.uk, NOT to the newsgroup mailing addresses. As usual, include the text in the body of the message as text on the Subject: line is ignored. To unsubscribe from all the lists at the UK node, use unsub bionet-news Please note that if the address in the list is different than the one in your mail message header, you will not be able to unsubscribe by this method. If you have problems, please mail biosci@daresbury.ac.uk. 4) The BIOSCI user address and research interest directory. ----------------------------------------------------------- Please take this opportunity to add your name, address, and research interest information to the BIOSCI User Address Database if you have not already done so. You can fill out the address form directly through our Web page at URL http://www.bio.net/adrform.html. The address database is reindexed nightly for WWW access (the URL is http://www.bio.net/). If you are not directly on the Internet but can reach it by e-mail, please use our waismail server to access the user directory. waismail use is described above. You can also request a user address form by e-mail from biosci-help@net.bio.net. Please check your database entry from time-to-time to see if your address information is still up-to-date. Because of our limited personnel resources, we ask that you resubmit a *complete* form to revise your entry; we only replace complete entries and do not have resources to edit old forms. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-fluorescent@net.bio.net Tue Dec 10 22:00:00 1996 Path: biosci!daresbury!not-for-mail From: Xavier Gansel Newsgroups: bionet.molbio.proteins.fluorescent Subject: GFP isforms in A. thaliana Date: 9 Dec 1996 12:11:59 -0000 Lines: 38 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <58gvmf$du7@mserv1.dl.ac.uk> Original-To: fluorpro@dl.ac.uk Dear Netters, Here is a mail to ask for advices on the use of GFP in A. thaliana. I wish to use GFP as a reporter gene to follow the regulation (activation/repression, tissue and cell expression, etc. ....) of a promoter. So, I want to get transgenic A.t. plants carrying a GFP under the control of this promoter. From the bibliography, it appears that their is a growing number of "plant optimised" GFP. The optimisations beeing on different aspects. Some of them are the optimisation of codon usage, the removal of a cryptic A.t. intron, the addition of a plant intron, mutations that shift light emission or enhance thermoresistance, anchoring in the ER to reduce toxicity of nuclear GFP accmlulation (Chui, 1996, Curr. Biol., 6, 325-330 // Davis, Weeds World Volume 3(ii) // Haseloff, TIG, 1951, 11, 328-329 // Haseloff, poster S44, 7th International Conference on Arabidopsis Research // Pang, Plant Physiol., 1996, 112, 893-900). But all those mutations/improvements are not present on the same plant-GFP. My questions are the following : 1/ on today statut, which version of plant improved GFP would you recommend to be use as the best reporter system in A.t. ? (the promoter to be studied may be weak and so require a very sensitive reporter gene to detect it) 2/ were could I get this "super" plant-GFP ? Of course any other advices (GFP version, detection protocols, etc ...) are welcome. Thanks for your help in this work, Your sincerely, Xavier GANSEL. From owner-fluorescent@net.bio.net Sat Dec 14 22:00:00 1996 Path: biosci!CMB.BIOSCI.WAYNE.EDU!hivlab From: hivlab@CMB.BIOSCI.WAYNE.EDU ("A. S. Goustin Lab") Newsgroups: bionet.molbio.proteins.fluorescent Subject: Re: Your GOD Loves You - YES YOU! Date: 15 Dec 1996 14:28:42 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 20 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <1212199600102165035202419509Someone@home.thinking.about.you.com> NNTP-Posting-Host: net.bio.net On Thu, 12 Dec 1996, Someone wrote: > I hope that you know your GOD loves you, no matter what GOD you believe in. > Thank your GOD for life and ask your GOD to let you live as your GOD would want > you to. When we look closely at GOD we begin to realize that we all believe in the > same GOD, we may see GOD in different ways, but GOD will always be GOD. There > can only be one GOD, and that one GOD loves us all, and wants nothing but the > best for us. If your in doubt, just remember GOD works in mysterious ways, there is > a reason for everything... > > God thank you. Should read "God thanks you", shouldn't it? Ominiscient, omnipotent, but grammatically incorrect. > > Someone > > > From owner-fluorescent@net.bio.net Sun Dec 15 22:00:00 1996 Path: biosci!nmu.edu!jrebers From: jrebers@nmu.edu (John Rebers) Newsgroups: bionet.molbio.proteins.fluorescent Subject: Drosophila cuticle autofluorescence Date: 16 Dec 1996 15:52:48 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 21 Sender: daemon@net.bio.net Distribution: world Message-ID: <2.2.16.19961216185306.3567bc0a@pop.mail.nmu.edu> NNTP-Posting-Host: net.bio.net I'm interested in using GFP as a reporter to study expression of a Manduca cuticle gene, by transfecting Drosophila with a Manduca cuticle promoter/GFP fusion, and then examining expression of the GFP in epidermal tissues. My concern is that Drosophila cuticle has a yellow-green autofluorescence. How hard is this to distinguish from GFP fluorescence? Would something like one of the blue-fluorescent proteins work, or would it best to use beta-galactosidase or another gene as the reporter? Thanks for any advice. John Rebers Department of Biology 1401 Presque Isle Avenues Northern Michigan University Marquette, MI 49855 906-227-1585 (office) 906-228-3617 (home) 906-227-2013 (FAX) e-mail address: jrebers@nmu.edu Note: do not send mail to jrebers@pop.mail.nmu.edu or to jrebers@VM.NMU.EDU - these may appear in the header, but will not work for my address. Thanks! From owner-fluorescent@net.bio.net Tue Dec 17 22:00:00 1996 Path: biosci!physiol.ox.ac.uk!tim.pragnell From: tim.pragnell@physiol.ox.ac.uk (Tim Pragnell Alias) Newsgroups: bionet.molbio.proteins.fluorescent Subject: (none) Date: 18 Dec 1996 04:05:10 -0800 Organization: University Laboratory of Physiology, Oxford Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: <32B7DDC5.554A@physiol.ox.ac.uk> NNTP-Posting-Host: net.bio.net info ukinfo From owner-fluorescent@net.bio.net Tue Dec 17 22:00:00 1996 Path: biosci!MAC.MB.WAU.NL!jan.carette From: jan.carette@MAC.MB.WAU.NL (peter) Newsgroups: bionet.molbio.proteins.fluorescent Subject: m-gfp5-ER versus EGFP Date: 18 Dec 1996 06:42:36 -0800 Organization: Agricultural University Wageningen Lines: 11 Sender: daemon@net.bio.net Distribution: world Message-ID: <32B810CE.63A0@mac.mb.wau.nl> Reply-To: jan.carette@MAC.MB.WAU.NL NNTP-Posting-Host: net.bio.net I would like to know wheter there are any studies where the m-gfp5-ER (Haseloff) is compared to the EGFP (Clontech; contains S65Tand Phe64-->Leu mutations combined with preferred human codon usage) with regard to the intensity of fluorescence, preferably in plant systems? Best regards, Jan Carette dept. Moleculair biology Agricultural University of Wageningen The Netherlands