From owner-pseudomonas@net.bio.net Fri Sep 08 23:00:00 1995 Path: biosci!agate!msunews!netnews.upenn.edu!dolphin.upenn.edu!lfmuscar From: lfmuscar@dolphin.upenn.edu (Lawrence Muscarella) Newsgroups: bionet.organisms.pseudomonas Subject: pseudo and UV light Date: 9 Sep 1995 18:42:38 GMT Organization: University of Pennsylvania Lines: 11 Message-ID: <42sn6u$kq7@netnews.upenn.edu> NNTP-Posting-Host: dolphin.upenn.edu X-Newsreader: TIN [version 1.2 PL2-upenn1.3] I am trying to get info on the use of UV light to eradicate the pseudomonas organism. DOes it work? DO UV lights prevent the growth of pseudomonas aeruginosa (sic.) is a poorly ventilated and moist room. Please provide me with any info on this matter. I would greatly appreciate the references of any papers that address this topic. Regards, Larry My E-mail address is: lfmuscar@alumni.upenn.edu From owner-pseudomonas@net.bio.net Fri Sep 08 23:00:00 1995 Path: biosci!daresbury!not-for-mail From: BIOSCI Administrator Newsgroups: bionet.organisms.pseudomonas Subject: test of pseudomo@daresbury.ac.uk Date: 9 Sep 1995 01:47:33 +0100 Lines: 1 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <42qo75$7dl@mserv1.dl.ac.uk> Reply-To: biosci-help@net.bio.net Original-To: pseudomo@dl.ac.uk test of pseudomo@daresbury.ac.uk From owner-pseudomonas@net.bio.net Fri Sep 08 23:00:00 1995 Path: biosci!NET.BIO.NET!biosci-help From: biosci-help@NET.BIO.NET (BIOSCI Administrator) Newsgroups: bionet.organisms.pseudomonas Subject: test of pseudomo@net.bio.net Date: 8 Sep 1995 17:48:56 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: Reply-To: biosci-help@net.bio.net NNTP-Posting-Host: net.bio.net test of pseudomo@net.bio.net From owner-pseudomonas@net.bio.net Fri Sep 08 23:00:00 1995 Path: biosci!biosci!not-for-mail From: kristoff@net.bio.net (David Kristofferson) Newsgroups: bionet.organisms.pseudomonas Subject: test of bionet.organisms.pseudomonas Date: 8 Sep 1995 17:21:51 -0700 Organization: BIOSCI International Newsgroups for Biology Lines: 11 Distribution: world Message-ID: <42qmmv$8d6@net.bio.net> NNTP-Posting-Host: net.bio.net This group is not yet ready for use. Please refrain from posting until an official opening announcement is made here. Thanks! Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-pseudomonas@net.bio.net Sat Sep 09 23:00:00 1995 Path: biosci!agate!newsxfer.itd.umich.edu!newsfeed.internetmci.com!howland.reston.ans.net!tank.news.pipex.net!pipex!in1.uu.net!zib-berlin.de!fu-berlin.de!news.dfn.de!uni-muenster.de!asterix.uni-muenster.de!katz From: katz@asterix.uni-muenster.de (Raphael Katz) Newsgroups: bionet.organisms.pseudomonas Subject: human telomerase Date: 10 Sep 1995 16:57:33 GMT Organization: Westfaelische Wilhelms-Universitaet Muenster, Germany Lines: 19 Message-ID: <42v5dt$1ib0@majestix.uni-muenster.de> NNTP-Posting-Host: asterix.uni-muenster.de X-Newsreader: TIN [version 1.2 PL2] Hallo, Wer verkauft menschliche Telomerase ? Welches Zellabor kann nach Auftrag Telomerase herstellen, z.B. durch Extraktion aus maligne entarteten Zellen gem„á Counter, Christopher M. et al., (1994) Proc. Natl. Acad. Sci. USA 91, 2900-2904 ? Who sells human telomerase, the telomere terminal transferase enzyme ? Who knows a cell laboratory, which can extract telomerase from carcinoma cells according Counter, Christo- pher M. et al., 1994 Proc. Natl. Acad. Sci. USA 91, 2900-2904 ? bye RAF From owner-pseudomonas@net.bio.net Sun Sep 10 23:00:00 1995 Path: biosci!biosci!not-for-mail From: biohelp@net.bio.net (BIOSCI Administrator) Newsgroups: bionet.organisms.pseudomonas Subject: PSEUDOMONADS/bionet.organisms.pseudomonas is ready! Date: 11 Sep 1995 11:39:46 -0700 Organization: BIOSCI International Newsgroups for Biology Lines: 194 Distribution: world Message-ID: <431vpi$ehi@net.bio.net> NNTP-Posting-Host: net.bio.net The PSEUDOMONADS/bionet.organisms.pseudomonas newsgroup is ready for operation. Please save these usage instructions for future reference!! PLEASE NOTE that many USENET sites do not allow automatic creation of new USENET groups!!! 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You can also access these same files via Gopher if you start a gopher session using net.bio.net as your gopher server. Gopher also allows you to view the individual messages within each monthly archive file. The files are in the PSEUDOMONADS directory. Postings to bionet.organisms.pseudomonas are also WAIS indexed and can be searched via either gopher or WAIS at our site. In gopher the option at net.bio.net is "Search Bionet USENET Articles" and in WAIS one should use the WAIS source biosci.src. This is a WAIS index of all BIOSCI/bionet messages including this newsgroup. Please see the BIOSCI FAQ for details. The FAQ can be requested from biosci-help@net.bio.net. Once again, if you have any administrative questions that require personal assistance, please address them to biosci-help@net.bio.net in the U.S. or biosci@daresbury.ac.uk in the UK. Best wishes for a successful newsgroup! Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-pseudomonas@net.bio.net Fri Sep 15 23:00:00 1995 Path: biosci!agate!msunews!uwm.edu!cs.utexas.edu!usc!howland.reston.ans.net!newsfeed.internetmci.com!usenet.eel.ufl.edu!usenet.cis.ufl.edu!usenet.ufl.edu!usenet From: "Marcus N. Morgan" Newsgroups: bionet.organisms.pseudomonas Subject: testing Date: 15 Sep 1995 14:29:26 GMT Organization: NERDC Lines: 2 Message-ID: <43c2k6$npj@no-names.nerdc.ufl.edu> NNTP-Posting-Host: morgan.nerdc.ufl.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (X11; I; SunOS 4.1.4 sun4m) X-URL: news:bionet.organisms.pseudomonas testing. first appearance at uf. From owner-pseudomonas@net.bio.net Fri Sep 15 23:00:00 1995 Path: biosci!agate!news.duke.edu!news-server.ncren.net!hearst.acc.Virginia.EDU!uunet!in1.uu.net!comp.vuw.ac.nz!canterbury.ac.nz!cantva!botn078 From: botn078@csc.canterbury.ac.nz Newsgroups: bionet.organisms.pseudomonas Subject: Cloning vectors... Date: 14 Sep 95 14:54:20 +1200 Organization: University of Canterbury, Christchurch, New Zealand Lines: 14 Message-ID: <1995Sep14.145420@cantva> NNTP-Posting-Host: cantva.canterbury.ac.nz Hi there. Can anyone out there advise me on cloning vectors? I work on a P. aureofaciens strain, but have difficulty due to most vectors being unstable in this background. Currently I am forced to use pLAFR3 for my work, and this is less than satisfactory! Any help would be great. Cheers, Mark Silby Genetics Lab Dept of Plant and Microbial Sciences University of Canterbury Christchurch, New Zealand. From owner-pseudomonas@net.bio.net Mon Sep 18 23:00:00 1995 Path: biosci!SUNFLOWER.BIO.INDIANA.EDU!cbauer From: cbauer@SUNFLOWER.BIO.INDIANA.EDU Newsgroups: bionet.organisms.pseudomonas Subject: PRE-DOCTORAL FELLOWSHIPS Date: 19 Sep 1995 11:37:36 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 52 Sender: daemon@net.bio.net Distribution: world Message-ID: <9509191833.AA09096@sunflower.bio.indiana.edu> NNTP-Posting-Host: net.bio.net ******************* PRE-DOCTORAL FELLOWSHIPS *********************** ******************* IN PLANT MOLECULAR BIOLOGY ********************** The Department of Biology at Indiana University-Bloomington has three predoctoral fellowships available to work in plant molecular biology laboratories. These fellowships are funded in part by the USDA National Needs Plant Biotechnology Fellowship program, and provide a stipend of $17,000 per year + tuition, health insurance and fees, for a total support of $27,000 per year. Fellowship recipients must be citizens or permanent residents of the U.S. The fellowship awardees can choose to work in any of the following laboratories: Carl Bauer: Genetic analysis of chlorophyll biosynthesis; regulation of photosynthesis gene expression by light and oxygen; phototaxis Mark Estelle: Genetic and molecular analysis of hormone action in Arabidopsis. Roger Hangarter: Plant physiology, the interaction of light, gravity, and hormones in regulating plant development. Roger Innes: Molecular genetics of plant-pathogen interactions. C. Cheng Kao: Mechanisms of plant-virus, and plant-bacteria interactions. Jeffrey Palmer: Molecular evolution; Origin and evolution of introns and plant organelle genomes. Robert Togasaki: Carbon assimilation and responses to oxidative stress in Chlamydomonas reinhardtii Mimi Zolan: Meiosis and DNA repair; Fungal genetics and genome evolution. More information regarding the research interests of the faculty, Ph. D degree programs and an application to the Department of Biology can be obtained at our web site ****http://www.bio.indiana.edu/**** or by writing to: Gretchen Clearwater, Administrative Assistant National Needs Fellowships Program Department of Biology Jordan Hall 138 Indiana University Bloomington, IN 47405 From owner-pseudomonas@net.bio.net Wed Sep 20 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!news.sprintlink.net!cs.utexas.edu!convex!news.onramp.net!news.tcst.com!dildog.lgc.com!news.sesqui.net!oitnews.harvard.edu!NewsWatcher!user From: mjcoyne@warren.med.harvard.edu (Michael Coyne) Newsgroups: bionet.organisms.pseudomonas Subject: Re: Cloning vectors... Date: 15 Sep 1995 21:20:14 GMT Organization: Channing Lab Lines: 34 Message-ID: References: <1995Sep14.145420@cantva> NNTP-Posting-Host: 134.174.147.40 In article <1995Sep14.145420@cantva>, botn078@csc.canterbury.ac.nz wrote: > Hi there. Can anyone out there advise me on cloning vectors? I work on a P. > aureofaciens strain, but have difficulty due to most vectors being unstable in > this background. Currently I am forced to use pLAFR3 for my work, and this is > less than satisfactory! > > Any help would be great. > > Cheers, > > Mark Silby > Genetics Lab > Dept of Plant and Microbial Sciences > University of Canterbury > Christchurch, New Zealand. Mark - You might try the pUCP plasmids - pUCP18 and pUCP19. These plasmids are dervived from pUC18 and pUC19 and, therefore, are high copy number and small - two additional pluses over the pLAFR plasmids. See Schweizer, H. P. 1991. Escherichia-Pseudomonas shuttle vectors derived from pUC18/19. Gene. 97:109-112. There is also a more recent paper in which Schweizer reports the sequence of the "1.8 kb Pst I stabilizing fragment of pRO1614" which was used to modify the pUC plasmids. I don't have the reference handy, sorry.... Can anyone help? Mike Coyne Channing Laboratory Harvard Medical School/Brigham & Women's Hospital Boston, MA From owner-pseudomonas@net.bio.net Thu Sep 21 23:00:00 1995 Path: biosci!agate!newsxfer.itd.umich.edu!tank.news.pipex.net!pipex!news.uoregon.edu!news.u.washington.edu!uw-beaver!cornellcs!travelers.mail.cornell.edu!newsstand.cit.cornell.edu!NewsWatcher!user From: tpd4@cornell.edu (Terry Delaney) Newsgroups: bionet.organisms.pseudomonas Subject: Re: Cloning vectors... Date: Thu, 21 Sep 1995 10:23:26 -0500 Organization: Cornell University Lines: 72 Sender: tpd4@cornell.edu (Verified) Message-ID: References: <1995Sep14.145420@cantva> NNTP-Posting-Host: 132 In article , mjcoyne@warren.med.harvard.edu (Michael Coyne) wrote: > In article <1995Sep14.145420@cantva>, botn078@csc.canterbury.ac.nz wrote: > > > Hi there. Can anyone out there advise me on cloning vectors? I work on a P. > > aureofaciens strain, but have difficulty due to most vectors being unstable in > > this background. Currently I am forced to use pLAFR3 for my work, and this is > > less than satisfactory! > > > > Any help would be great. > > > > Cheers, > > > > Mark Silby > > Genetics Lab > > Dept of Plant and Microbial Sciences > > University of Canterbury > > Christchurch, New Zealand. > > Mark - > > You might try the pUCP plasmids - pUCP18 and pUCP19. These plasmids are > dervived from pUC18 and pUC19 and, therefore, are high copy number and > small - two additional pluses over the pLAFR plasmids. See Schweizer, H. > P. 1991. Escherichia-Pseudomonas shuttle vectors derived from pUC18/19. > Gene. 97:109-112. > > There is also a more recent paper in which Schweizer reports the sequence > of the "1.8 kb Pst I stabilizing fragment of pRO1614" which was used to > modify the pUC plasmids. I don't have the reference handy, sorry.... Can > anyone help? > > Mike Coyne > Channing Laboratory > Harvard Medical School/Brigham & Women's Hospital > Boston, MA The reference is: Construction of improved Escherichia-Pseudomonas shuttle vectors derived from pUC18/19 and sequence of the region required for their replication in Pseudomonas aeruginosa. West SE; Schweizer HP; Dall C; Sample AK; Runyen-Janecky LJ Gene (NETHERLANDS) Oct 11 1994, 148 (1) p81-6, ISSN 0378-1119 The nucleotide sequence of the 1.9-kb PstI fragment from pRO1614, that allows stable maintenance of pMB1 (ColE1)-based cloning vectors in Pseudomonas, was determined. This fragment encodes a putative origin of replication (ori), a replication-controlling protein, and the C terminus of the Tn3 beta-lactamase-encoding gene. Improved versions of the broad-host-range plasmid vectors, pUCP18 and pUCP19, were constructed by deletion of nonessential DNA or replacement of nonessential DNA with an antibiotic-resistance cassette. Cheers, Terry Delaney mwwmwwmwwmwwmwwmwwmwwmwwmwwmwwmwwm Terrence P. Delaney Department of Plant Pathology Cornell University 334 Plant Science Building Ithaca, NY 14853-4203 (607) 255-7856 (607) 255-4471 (fax) tpd4@cornell.edu mwwmwwmwwmwwmwwmwwmwwmwwmwwmwwmwwm From owner-pseudomonas@net.bio.net Thu Sep 21 23:00:00 1995 Path: biosci!agate!msunews!uwm.edu!psuvax1!gvls1!newsfeed.pitt.edu!uunet!in2.uu.net!usenet.eel.ufl.edu!warwick!bham!bcs177.bham.ac.uk!J.D.Brinck From: J.D.Brinck@bham.ac.uk (Jason Brinck) Newsgroups: bionet.organisms.pseudomonas Subject: Problems with Pseudomonas putida miniprep Date: Thu, 21 Sep 1995 11:58:54 Organization: The University of Birmingham Lines: 41 Message-ID: NNTP-Posting-Host: bcs177.bham.ac.uk X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] Hi all, Does anyone have any good ideas to help me get round the following problem: I routinely grow a strain of Pseudomonas putida, which contains a recombinant plasmid, as a continuous culture. I am trying to analyse plasmid DNA isolated from samples taken at different time points during the experiment. As well as observing if the size of the plasmid remains unchanged I would also like to see if the intracellular concentration of plasmid varies. In an attempt to do this I have harvested equal amounts of cells at each sample (the equivalent of 1ml of O.D. 2 culture) which I then store at -20 C as a dry pellet. At the end of the experiment plasmid DNA is isolated from all the samples using a standard miniprep protocol. The problem I have encountered is a complete lack of consistancy / reproducability. The amount of plasmid DNA I recover from minipreps is highly variable, even between duplicate samples. Also, the quality of DNA obtained is often poor such that when I try to linearise the plasmid with HindIII a lot of the samples only partly cut. I have tried different miniprep methods (alkaline lysis, boiling) as well as kits (Bio101 RPM kit) but seen little improvement. I know that this strain is not very "user-friendly". It has a low viability on plates (up to a week) so I was wondering if part of the problem is how I store the samples until I miniprep them all at the end of the experiment. Might storing them at -70 C improve things. Other than that all I can think of is storing the sample as a glycerol but I think that would then cause problems when I come to miniprep the samples. Even so, I'm still not happy with the miniprep method as I still get uncut bands when I miniprep fresh culture and linearise it. So, any suggestions would be greatly appreciated as time is running out for me! Jason Brinck, School of Biochemistry Birmingham University, UK From owner-pseudomonas@net.bio.net Fri Sep 22 23:00:00 1995 Path: biosci!bloom-beacon.mit.edu!newsfeed.internetmci.com!usenet.eel.ufl.edu!usenet.cis.ufl.edu!usenet.ufl.edu!gnv.ifas.ufl.edu!icbr.ifas.ufl.edu!bwr Newsgroups: bionet.organisms.pseudomonas Subject: test-ignore Message-ID: <1995Sep22.100045.1@icbr.ifas.ufl.edu> From: bwr@icbr.ifas.ufl.edu Date: 22 Sep 95 10:00:45 -0500 Organization: ICBR Nntp-Posting-Host: icbr.ifas.ufl.edu Lines: 1 test-ignore From owner-pseudomonas@net.bio.net Sat Sep 23 23:00:00 1995 Path: biosci!agate!dog.ee.lbl.gov!news.cs.utah.edu!cs.utexas.edu!news.sprintlink.net!tank.news.pipex.net!pipex!usenet.eel.ufl.edu!usenet.cis.ufl.edu!usenet.ufl.edu!usenet From: "Bruce W. Ritchings" Newsgroups: bionet.organisms.pseudomonas Subject: Re: Problems with Pseudomonas putida miniprep Date: 23 Sep 1995 18:44:56 GMT Organization: University of Florida Lines: 8 Message-ID: <441kj9$9v0@no-names.nerdc.ufl.edu> References: NNTP-Posting-Host: bruce.med.ufl.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.2b5 (Windows; I; 16bit) Jason, when you do the alkaline lysis minipreps, are you washing the cells with STE before continuing on with solution I? If not, that's a first place to start (see Sambrook et al Cloning Manual, p.1.25). My guess would be that storing the cultures at any freezing temperature would not be the best idea--as soon as the broken cells are warmed up, the nucleases would become active. Any chance you can carry the miniprep samples through at least to the ethanol stage before storing them?