From owner-7tms_r@net.bio.net Mon Jul 06 23:00:00 1998 Path: biosci!biosci!not-for-mail From: IARC p53 Database Newsgroups: bionet.molbio.proteins.7tms_r Subject: IARC p53 database update Date: 7 Jul 1998 12:15:53 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 29 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6nts59$t61@net.bio.net> NNTP-Posting-Host: net.bio.net Dear All, The International Agency for Research on Cancer is pleased to announce the July release of the p53 mutation database which covers somatic mutations in human tumors and cell lines. Please have a look at our home page http://www.iarc.fr/p53/homepage.htm where you can directly access the p53 flat file, the SRS version of the database and an applet viewer of the p53 mutations. The current update contains 9378 mutations which represent 999 different references. Any comments or suggestions??? We would be greatful to hear from you. Tina Hernandez International Agency of Research on Cancer IARC p53 Mutation Database 150 Cours Ablert Thomas Lyon 69372 France http://www.iarc.fr/p53/Homepage.htm Tel: 33 (0)4 72 73 84 85 Fax: 33 (0)4 72 73 85 75 http://www.iarc.fr/p53/Homepage.htm From owner-7tms_r@net.bio.net Mon Jul 06 23:00:00 1998 Path: biosci!biosci!not-for-mail From: "Steve Qi" Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: reporter gene for IP3 Date: 7 Jul 1998 12:15:56 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 74 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6nts5c$t6j@net.bio.net> References: <6dmbeq$4hs@net.bio.net> NNTP-Posting-Host: net.bio.net Steve- NFAT can be a pretty good reporter if NFAT is expressed in your cells. PLC activation is necessary and sufficient for NFAT mediated transcription so it is probably one of the truest reporters for PLC activity, I mean, everything including sand can induce AP1-mediated transcription so that is not very specific. If NFAT is not expressed in your cells, it won't work well, so you really need a good cell line. PC12 cells express it and it would function well in there (Boss, V., Joshi-Talpade, D. and T.J. Murphy. Induction of NFAT-mediated transcription by Gq-protein coupled receptors in lymphoid and non-lymphoid cells, J. Biological Chem. 271:10429-10432 (1996).) Another key to a good NFAT response to an agonist is also in the strength of capacitive Ca entry in the cells. Agonists that spike Ca, rather than giving a sustained Ca elevation will not always stimulate NFAT -transcription real well. A general strategy in making more sensitive reporters that is worth trying is to use a minimal promoter from a highly inducible gene. By minimal promoter, I mean one in which all enhancers have been deleted so it serves only as an RNAPolII docking site. Typically, you want a well-behaved TATA box promoter, rather than a promoter with an INR or some such. We use the minimal IL-2 promoter from the human gene (-72 to +48). Basal activity from this is virtually unmeasurable in our hands in several cells using luciferase. Then, link up concatemers of a response element (such as the NFAT RE) upstream of that. Try to keep the monomers in the repeat to ~20-30 bp, and the whole concatemer ~60-120 bp in size. We found this strategy works much better than an uninducible minimal promoter, such as that from the TK or CMV genes, which a lot of people use. My best guess for why your construct doesn't work is that you've got quite a few weak enhancers in the ~550 bp, that sum to give you high basal. The phorbol response element likely works fine, but the background noise is too high for it appear robust. Steve Qi wrote: > Dear fellow colleagues, > I am currently trying to construct a reporter gene cell line for monitoring > the IP3/DAG signal pathway. I have cloned the ICAM-1 promoter and inserted > the 5' untranslated region (-616 to -51) as a XhoI - SalI fragment into > pGL3basic (Promega). The construct response to PdBU (5 - 7X) but has very > high basal activity. Can anybody give me some suggestions on how to reduce > the background or suggest another response element? > Thanks > Steve (sqi@ferring.demon.co.uk) - -- T.J. Murphy, Ph.D. 404-727-2467 Assistant Professor 404-727-0365 (fax) Department of Pharmacology tmurphy@pharm.emory.edu Emory University School of Medicine 5031 O.W. Rollins Research Building Atlanta, GA 30322 http://www.emory.edu/PHARMACOLOGY/MURPHY/ "If we're not driving paradigms, then we're out driving Titleists" From owner-7tms_r@net.bio.net Mon Jul 06 23:00:00 1998 Path: biosci!biosci!not-for-mail From: "Steve Qi" Newsgroups: bionet.molbio.proteins.7tms_r Subject: ip3 reporter Date: 7 Jul 1998 12:16:21 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 22 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6nts65$t8v@net.bio.net> NNTP-Posting-Host: net.bio.net Steve; I have no suggestions, but would love to have the construct once the bugs are worked out! MAL Michael A. Levine, M.D. Johns Hopkins University School of Medicine Division of Endocrinology and Metabolism 863 Ross Research Building 720 Rutland Avenue Baltimore, MD 21205 T 410.955.7233 F 410.955.0841 From owner-7tms_r@net.bio.net Mon Jul 06 23:00:00 1998 Path: biosci!biosci!not-for-mail From: Athan Kuliopulos Newsgroups: bionet.molbio.proteins.7tms_r Subject: Two Signal Transduction Postdoc Positions Avail-Boston Date: 7 Jul 1998 12:18:12 -0700 Organization: NEMC Lines: 59 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6nts9k$mu@net.bio.net> Reply-To: "Kuliopoulos@HemOnc"@NEMC.nemc.org NNTP-Posting-Host: net.bio.net Two Postdoctoral Positions Available Molecular Cardiology Research Institute Tufts University School of Medicine-NEMC Boston, MA We have an immediate opening for two Postdoctoral Fellows to work in the field of molecular signaling and peptide-protein recognition. The projects focus on the human thrombin receptor. The thrombin receptor is activated by thrombin cleavage of the receptor exodomain and exposure of an N-terminal tethered ligand that binds to the body of the receptor. Receptor activation precipitates complex signaling events culminating in platelet aggregation, wound healing, and cellular proliferation. Since chronic activation of the receptor may lead to coronary artery disease, stroke, and other vascular diseases, preventing thrombin receptor activation is of pharmacologic interest. 1) Macromolecular Structural Studies of the Resting and Activated States of Thrombin Receptor Extracellular Domains. NMR structural studies of the thrombin receptor exodomain in activated and resting forms are currently in progress and a preliminary structure has been generated for the activated exodomain. Future projects include solving the structure of the exodomain complexed with extracellular loops. Insight into the molecular interactions between the exodomain and the body of the receptor should provide leads for the development of novel anti-thrombin receptor agents in collaboration with a pharmaceutical company. The NMR facility is located in the Medical School Biochemistry Department and current instrumentation include a new Bruker 600 MHz and updated 500 MHz magnets along with several SGI workstations. Our lab has close collaborations with NMR spectroscopists who provide additional technical expertise. 2) Thrombin-Cell Surface Protein Interactions; Development of Cell Surface-Specific Anti-Thrombotic Agents. During coagulation, the physiologic concentration of thrombin exceeds that of its thrombin receptor substrate. Therefore, thrombin has a difficult task of discriminating among the various cell surface proteins to find the receptor and cleave it in the millisecond time range. We are interested in exploring this unusual mechanism of substrate-assisted domain cleavage by thrombin and using this information to develop a novel class of cell surface anti-thrombin agents. The laboratory is located within the Molecular Cardiology Research Institute, a modern, state-of-the-art facility with a staff of 40 investigators including technical support. Qualifications for this position are a Ph.D. degree. Candidates with training in NMR who would like to acquire expertise in molecular biology are encouraged to apply. Interested candidates should e-mail a description of their research interests, a CV, and names of three references to: Athan Kuliopulos, MD., Ph.D. Assistant Professor of Medicine and Biochemistry Molecular Cardiology Research Institute Tufts-NEMC Box 832 750 Washington Street Boston, MA 02111 617-636-8482 617-636-4833 (fax) akuliopu@opal.tufts.edu From owner-7tms_r@net.bio.net Mon Jul 06 23:00:00 1998 Path: biosci!biosci!not-for-mail From: "Steve Qi" Newsgroups: bionet.molbio.proteins.7tms_r Subject: IP3 reporter gene Date: 7 Jul 1998 12:16:25 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 16 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6nts69$3u@net.bio.net> NNTP-Posting-Host: net.bio.net Steve, Are you familiar with prior published work by C. Stratowa, 1995, J. of Receptor and Signal Transduction Research 15: 617-630? In this study, they constructed cell lines expressing individual human neurokinin receptor subtypes stably transformed with the luciferase gene under the control of a promoter containing TPA response elements. We might be interested in obtaining such a line from you, depending upon its characteristics, or alternatively collaborating to establish a novel line in a particular host cell. Are either of these options of possible interest to you? Jeffrey Herz, Ph.D. Director of Pharmacology From owner-7tms_r@net.bio.net Mon Jul 06 23:00:00 1998 Path: biosci!biosci!not-for-mail From: "Steve Qi" Newsgroups: bionet.molbio.proteins.7tms_r Subject: RE: reporter gene for IP3 Date: 7 Jul 1998 12:16:23 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 37 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6nts67$t9b@net.bio.net> NNTP-Posting-Host: net.bio.net Created on 03/05/98 8:00 AM Attention: Steve I am not able to offer direct advice/help. I am curious, however, what cell line you are using and how you determine the control basal activity of the pathway. I have studied this pathway in a number of cells and I often find very high IP3 production in cells we have not stimulated at all. What do you expect PKC activation by phorbol esters to do in this system? Best regards, Iain L. O. Buxton Professor of Pharmacology Phone: (702)784-6956 (Dept.); 784-1566(Office); 784-4120 (Lab) University of Nevada FAX: (702)784-1620 (Dept.); 784-1378(Office) School of Medicine e-mail: buxton@med.unr.edu (Office) buxton@aci.net (Home) Howard Medical Sciences Bldg. Pager: (702)858-4123 Laboratory Suite 216 Reno, NV 89557 - -----Original Message----- From: Steve Qi [SMTP:sqi@ferring.demon.co.uk] Sent: Wednesday, March 04, 1998 7:08 AM To: nobody@net.bio.net Subject: reporter gene for IP3 Dear fellow colleagues, I am currently trying to construct a reporter gene cell line for monitoring the IP3/DAG signal pathway. I have cloned the ICAM-1 promoter and inserted the 5' untranslated region (-616 to -51) as a XhoI - SalI fragment into pGL3basic (Promega). The construct response to PdBU (5 - 7X) but has very high basal activity. Can anybody give me some suggestions on how to reduce the background or suggest another response element? Thanks Steve (sqi@ferring.demon.co.uk) From owner-7tms_r@net.bio.net Sun Jul 12 23:00:00 1998 Path: biosci!biosci!not-for-mail From: nevek@teleport.com (Kim Neve) Newsgroups: bionet.molbio.proteins.7tms_r Subject: G protein antibodies Date: 13 Jul 1998 13:40:37 -0700 Organization: Oregon Health Sciences University Lines: 4 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6odrc5$7ib@net.bio.net> NNTP-Posting-Host: net.bio.net Does anybody know of any (commercial) antibodies for transducin? How about an antibody that will recognize alpha-olf but not alpha-s? thanks, Kim Neve From owner-7tms_r@net.bio.net Sun Jul 12 23:00:00 1998 Path: biosci!biosci!not-for-mail From: nevek@teleport.com (Kim Neve) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: G protein antibodies Date: 13 Jul 1998 16:55:48 -0700 Organization: Oregon Health Sciences University Lines: 16 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6oe6q4$2gr@net.bio.net> References: <6odrc5$7ib@net.bio.net> NNTP-Posting-Host: net.bio.net nevek@teleport.com (Kim Neve) wrote: >Does anybody know of any (commercial) antibodies for transducin? How >about an antibody that will recognize alpha-olf but not alpha-s? >thanks, >Kim Neve I am following up on my own message because I realized that I was not clear in the first message. It is not that I am too lazy to go check the catalogs - I am hoping that somebody can give me information about antibodies that s/he has used successfully, preferably for western blotting. Also, it does not matter if the antibody is _commercially_ available, just available. Thanks, Kim Neve From owner-7tms_r@net.bio.net Sun Jul 12 23:00:00 1998 Path: biosci!biosci!not-for-mail From: daemon@net.bio.net Newsgroups: bionet.molbio.proteins.7tms_r Subject: POSITION: Scientist, Tumor/Angiogenesis In Vitro Date: 13 Jul 1998 13:38:47 -0700 Organization: Sedona Internet Services, Inc. Lines: 35 Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6odr8n$7d3@net.bio.net> Reply-To: rathbun@sedona.net NNTP-Posting-Host: net.bio.net We are seeking a PhD-level scientist with experience in angiogenesis and/or tumor research for a Discovery Research position with our client biopharmaceutical company. He/She will conduct research examining the role of various factors in In Vitr systems and will coordinate external collaborations in this area. Qualifications: =B7 Ph.D in appropriate Biological science,=20 =B7 Experience in use of In Vitro models to study actions of various factors with the emphasis on: Endothelial Cell Assays, Proliferation, Survival, Migration, Tube Formation, Sprouting= , Extracellular Matrix Production =B7 Use of Retroviral systems for reporter gene expression =B7 Differential display techniques. If you have an interest in this or other opportunities, please send u= s your resume/CV as an Attached File to an email or send by mail/FAX to RS&A to the attention of Ann Rathbun, Managing Director. All correspondence is held in strict confidence. Rathbun, Sapir & Associates P.O. Box 2337 =20 Sedona, AZ 86339-2337 * USA (520) 203-0074 Office (520) 203-0075 FAX=20 E-mail: rathbun@sedona.net From owner-7tms_r@net.bio.net Sun Jul 12 23:00:00 1998 Path: biosci!biosci!not-for-mail From: BIOSCI Administrator Newsgroups: bionet.molbio.proteins.7tms_r Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 13 Jul 1998 13:38:07 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 232 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6odr7f$7c9@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. 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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. From owner-7tms_r@net.bio.net Sun Jul 12 23:00:00 1998 Path: biosci!biosci!not-for-mail From: nevek@teleport.com (Kim Neve) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Postdoctoral Position Date: 13 Jul 1998 13:40:32 -0700 Organization: Oregon Health Sciences University Lines: 9 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6odrc0$7ht@net.bio.net> NNTP-Posting-Host: net.bio.net A Postdoctoral Position is available immediately at Oregon Health Sciences University for work focusing on molecular pharmacological characterization of dopamine receptors and G proteins - see http://www.teleport.com/~nevek/. Experience in techniques of molecular biology, protein chemistry, or biochemical pharmacology is essential, as well as enthusiasm and ambition. Send CV and names of three references to: Dr. Kim Neve, VA Medical Center (R&D-30), 3710 SW US Veterans Hospital Road, Portland, OR 97201. FAX: 503-721-7839; Email: nevek@ohsu.edu. From owner-7tms_r@net.bio.net Sun Jul 12 23:00:00 1998 Path: biosci!biosci!not-for-mail From: rathbun@sedona.net Newsgroups: bionet.molbio.proteins.7tms_r Subject: POSITION: Product Manager/Molecular Biology Products Date: 13 Jul 1998 13:39:37 -0700 Organization: Sedona Internet Services, Inc. Lines: 86 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6odra9$7f4@net.bio.net> Reply-To: rathbun@sedona.net NNTP-Posting-Host: net.bio.net COMPANY SUMMARY: The company is mid-sized company specializing in the development and manufacture of innovative products serving the life science research market. The company currently offers more than 2,000 products designed to facilitate gene expression and analysis, gene cloning, PRC analysis and many other molecular biology techniques used in developmental genetics, cellular biology, cancer research, antisense DNA research, genome mapping, neurobiology, oncology and several other areas. The company is very technology oriented and places great emphasis on quality, innovation, fiscal stability and responsiveness and responsibility towards customers and the community. JOB SUMMARY: This Product Manager will be responsible for sales revenue and growth of specific product lines which may include any of the following: gene expression products, functional analysis products, protein expression products or general gene reporter products. Due to its considerable growth rate, the company offers substantial advancement for suitably qualified applicants.=20 PRIMARY RESPONSIBILITIES INCLUDE: =B7 Technology licensing. =B7 Produce line strategic development. =B7 Alliance building and maintenance. =B7 Market research. =B7 Competitor analysis. =B7 Sales/distributor training and support=20 =B7 Management of the transfer of new products from research and development to operations. =B7 Development of global launch platforms and product positioning strategies. =B7 Satisfying revenue growth targets and specific product lines. JOB REQUIREMENTS Education and experience: =20 =B7 BA(S), MA(S), or Ph.D. in molecular biology, biochemistry, cellular biology or virology from an accredited institution either domestic U.S. or foreign with preference given to candidates with molecular biology background. =B7 Business training or experience in product management with preference given to candidates with an MBA in a relevant discipline. =B7 3-5 years of technical experience in molecular biology with preference being given to candidates with directly-related work experience in companies which product molecular biology kits and reagents. =B7 Minimum of two years of directly-relevant product management experience with preference being given to candidates with product management experience both domestic and international in scope. Knowledge and Skills Required: =B7 Knowledge of modern molecular biology, cell biology and biochemical techniques and their use in both research and academic environments. =B7 Good leadership and communication skills along with good analytical skills are essential. =B7 Candidates with exceptional detail-orientation are particularly sought after. =B7 Must possess excellent communications skills to interface with scientists both within and without the organization. This includes excellent proficiency in English (both written and verbal) and in the use of the scientific literature =B7 Computer literacy to include operation of a Personal Computer (IBM clone and/or Mac); proficiency in the use of the Internet. =B7 Knowledge of and experience in GMP/GLP environments. =B7 Able to manage multiple priorities and deadlines in an expedient and decisive manner. =B7 Must have sufficient emotional maturity and fortitude to flourish in an environment with constantly changing workloads and work requirements and must evolve with the position. If you have an interest in this or other opportunities, please send us your resume/CV as an Attached File to an email or send by mail/FAX to RS&A to the attention of Ann Rathbun, Managing Director. All correspondence is held in strict confidence. Rathbun, Sapir & Associates P.O. Box 2337 =20 Sedona, AZ 86339-2337 * USA (520) 203-0074 Office (520) 203-0075 FAX=20 E-mail: rathbun@sedona.net From owner-7tms_r@net.bio.net Sun Jul 12 23:00:00 1998 Path: biosci!biosci!not-for-mail From: rathbun@sedona.net Newsgroups: bionet.molbio.proteins.7tms_r Subject: POSITION: Scientist, Tumor/Angiogenesis In Vivo Date: 13 Jul 1998 13:39:27 -0700 Organization: Sedona Internet Services, Inc. Lines: 34 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6odr9v$7ef@net.bio.net> Reply-To: rathbun@sedona.net NNTP-Posting-Host: net.bio.net We are seeking a PhD-level scientist with experience in angiogenesis and/or tumor research for a Discovery Research position with our client biopharmaceutical company. He/She will conduct research examining the role of various factors in In Vivo models and will coordinate external collaborations in this area. This scientist will be working with: =A7 Recombinant proteins that agonize or antagonize factors =A7 Transgenic models in which proteins are knocked-out or overexpressed resulting in vascular phenotypes Qualifications: =B7 Ph.D in appropriate Biological science=20 =B7 Experience in use of In Vivo models to study actions of various factors with the emphasis on: Tumor Models, Corneal Pocket Model, Matrigel Angiogenesis Model, CAM assay, Wound Healing Models, Ischemic Models, Use of Adenoviral and Retroviral Protein Expression in the above systems. If you have an interest in this or other opportunities, please send us your resume/CV as an Attached File to an email or send by mail/FAX to RS&A to the attention of Ann Rathbun, Managing Director. All correspondence is held in strict confidence. Rathbun, Sapir & Associates P.O. Box 2337 =20 Sedona, AZ 86339-2337 * USA (520) 203-0074 Office (520) 203-0075 FAX=20 E-mail: rathbun@sedona.net From owner-7tms_r@net.bio.net Sat Jul 18 23:00:00 1998 Path: biosci!biosci!not-for-mail From: g.s.stewart@bham.ac.uk (Grant Stewart) Newsgroups: bionet.molbio.proteins.7tms_r Subject: cell lysate problem Date: 19 Jul 1998 16:56:56 -0700 Organization: CRC Institute for Cancer Studies, Birmingham University Lines: 23 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6ou148$bgq@net.bio.net> NNTP-Posting-Host: net.bio.net When I am preparing samples for Western I do the following: Thaw pellet of gamma-radiation treated LCLs (~4,000,000 cells) at 37oC. Resuspend pellet in 200ul UTB buffer (9M urea, 150mM B-mercaptoethanol, 50mM Tris pH7.5). Sonicate for 2x15 seconds, keeping cool, then centrifuge for 25mins at 35000 rpm at 10oC. The problem is that after centrifugation I get this viscous bubbly layer floating on the top of the centrifuged lysate. There is a pellet therefore it has been centrifuged properly. Has anyone had this problem and do they know what the layer is? I don't think that it is unbroken DNA/histone as it has been thoroughly sonicated. The layer is quite viscous which suggests its protein but it is in 9M urea so it should have dissolved. If anyone can suggest what this is then can you email me at: g.s.stewart@bham.ac.uk Thanks. G.Stewart. From owner-7tms_r@net.bio.net Sat Jul 18 23:00:00 1998 Path: biosci!biosci!not-for-mail From: edvard@fagmed.uit.no (Oyvind Edvardsen) Newsgroups: bionet.molbio.proteins.7tms_r Subject: tinyGRAP query logging has been turned OFF Date: 19 Jul 1998 16:55:34 -0700 Organization: Univ. of Tromso, IMB Lines: 37 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6ou11m$bg1@net.bio.net> Reply-To: edvard@fagmed.uit.no NNTP-Posting-Host: net.bio.net Dear tinyGRAP mutant database users, As an aid to tackle problems with the most recent version of the=20 database I turned on logging of queries when releasing the most recent version of the database a couple of months ago. Fortunately, I've found virtually no problems and detailed query logging has now been turned OFF. The stored query log has now been removed and has never been backed up or printed. Furthermore, I discovered today that there was a problem with the field in the query form where you can search for author names, words from paper titles etc. In fact words from paper titles could not be located!! This has been fixed. The only non-volatile address to the database is as usual http://www-grap.fagmed.uit.no/GRAP/homepage.html Sincerely, =D8yvind Edvardsen - -o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-= o-o- ___ _ =20 | /| _| |_ _| __ _ _| _ _ |/_| \/ \/ | |\| |_| |_ |_| \/ |_|_ | |_| _\ |_' |\| / _________________________________________________________________________= ____ School of Medicine | Dept. of Pharmacology, IMB | TelePhone: +47 77 64 53 42 University of Troms=F8 | TeleFax: +47 77 64 53 10 MH, Breivika | Email: edvard@fagmed.uit.no N-9037 TROMS=D8, NORWAY | URL: http://atf1.fagmed.uit.no/mgl.= html - -------------------------------------------------------------------------= - ----- From owner-7tms_r@net.bio.net Sat Jul 18 23:00:00 1998 Path: biosci!biosci!not-for-mail From: "Carol B. Fowler" Newsgroups: bionet.molbio.proteins.7tms_r Subject: Muscarinic antibodies to extracellular loops Date: 19 Jul 1998 16:54:03 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6ou0ur$bfb@net.bio.net> NNTP-Posting-Host: net.bio.net Hi, I'm a graduate student working in Henry Moberg's lab at the University of Michigan. I'm currently trying to stain cells expressing human muscarinic (1 and 2) mutants and I have had little luck locating antibodies. Any information you could give me on antibodies raised to the extracellular loops of either receptor would be greatly appreciated. Thanks for your time Carol Fowler cbfowler@umich.edu