DROSOPHILA INFORMATION NEWSLETTER Volume 11, July 1993 The Drosophila Information Newsletter has been established with the hope of providing a timely forum for informal communication among Drosophila workers. The Newsletter will be published quarterly and distributed electronically, free of charge. We will try to strike a balance between maximizing the useful information included and keeping the format short; priority will be given to genetic and technical information. Brevity is essential. If a more lengthy communication is felt to be of value, the material should be summarized and an address made available for interested individuals to request more information. Submitted material will be edited for brevity and arranged into each issue. Research reports, lengthy items that cannot be effectively summarized, and material that requires illustration for clarity should be sent directly to Jim Thompson (THOMPSON@AARDVARK.UCS.UOKNOR.EDU) for publication in DIS. Materials appearing in the Newsletter will be reprinted in DIS. Back issues of DIN are available from FlyBase in the directory flybase/news or in News/ when accessing FlyBase with Gopher. Material appearing in the Newsletter may be cited unless specifically noted otherwise. Material for publication should be submitted by e-mail. Figures and photographs cannot be accepted at present. Send technical notes to Carl Thummel and all other material to Kathy Matthews. The e-mail format does not allow special characters to be included in the text. Both superscripts and subscripts have been enclosed in square brackets; the difference should be obvious by context. Bold face, italics, underlining, etc. cannot be retained. Please keep this in mind when preparing submissions. To maintain the original format when printing DIN, use Courier 10cpi font on a standard 8.5" x 11" page with 1" margins. Drosophila Information Newsletter is a trial effort that will only succeed if a broad segment of the community participates. If you have information that would be useful to your colleagues, please take the time to pass it along. The editors: Carl Thummel Kathy Matthews Dept. of Human Genetics Dept. of Biology Eccles Institute - Bldg. 533 Indiana University University of Utah Bloomington, IN 47405 Salt Lake City, UT 84112 812-855-5782; FAX/2577 801-581-2937; FAX/5374 MATTHEWK@INDIANA.EDU CTHUMMEL@HMBGMAIL.MED.UTAH.EDU MATTHEWK@INDIANA.BITNET *** To add your name to the Newsletter distribution list, send one of the following E-mail messages from the account at which you wish to receive DIN. Via Bitnet -- To: LISTSERV@IUBVM Subject: Message: SUB DIS-L Your real name Via Internet -- To: LISTSERV@IUBVM.UCS.INDIANA.EDU Subject: Message: SUB DIS-L Your real name LISTSERV will extract your user name and node from the E- mail header and add you to the list. Use your Internet address if you have one. You will receive confirmation by E-mail if you have successfully signed on to the list. If you are on the list and do not wish to receive DIN, or you want to remove a soon-to-be- defunct address, replace SUB in the above message with UNS. The SUB command can also be used to correct spelling errors in your real name; the new entry will simply replace the old as long as it was sent from the same USERID@NODE address. *** DIN Vol. 11 TABLE OF CONTENTS >Introduction to Drosophila Information Newsletter >How to subscribe to the Newsletter >TABLE OF CONTENTS >ANNOUNCEMENTS >1994 National Drosophila Conference >Recent additions to FlyBase >Bloomington Stock Center News >REQUESTS FOR MATERIALS >Inserts or lethals in 17AB >Df(3L)Mg >ms(2)E8 and ms(2)E9 >Documents for Drosophila thesaurus >DIS 1-33 >MATERIALS AVAILABLE >Chromosome 3 homozygous-by-descent lines >TECHNICAL NOTES >Double-sided sticky tape for embryo injections >Cryopreservation of Drosophila embryos >Materials from the Drosophila Genome Center >GENETIC NOTES >Updates and corrections to the Redbook *** ANNOUNCEMENTS DROSOPHILA CONFERENCE The 1994 National Drosophila Conference will be held April 20-24, 1994 in Chicago, Illinois. Victoria Finnerty, Emery University, is the meeting organizer. Send suggestions for the 1994 conference format to VICTORIA@BIOLOGY.EMORY.EDU. *** RECENT ADDITIONS TO FLYBASE *A major new FlyBase product has been released to the FlyBase server at Indiana, and is available either by Gopher (in Flybase/References) or by ftp (in flybase/refs directory). It is a unified list of publications concerning Drosophila. Drawn from a variety of sources (described in references-sources.txt) it includes 51552 entries, from 1684 to 1993. All are full references with as complete bibliographic information as is available. You can search this reference list via Gopher. The reference file is available in a few formats including refer, as readable by many bibliographic programs including Endnote, Pro-Cite and Refer, and comma-separated-values (csv) as used by many spreadsheet and database programs. It is impossible that this file is either 'complete' or free of errors. Mail flybase-refs@morgan.harvard.edu with additions and corrections. * The stock list of the National Drosophila Species Resource Center at Bowling Green is now available on FlyBase. Three versions of the list are posted: 1) species-center.rtf contains the center's catalogue in rich text format, which retains formatting information from the original word- processing file provided by the stock center. If you have a word processing program that handles rtf you can print a copy of the center's catalogue from this file; 2) species-center.txt is a database/spreadsheet format file; 3) species-center.rpt is a file in report format more easily read by humans than the txt file (when searching the stock list with gopher, information from this report file is returned). See the document species-center.doc for further information about the species stock center and these files. All of these files are in the directory flybase/stocks/stock- centers. *** BLOOMINGTON STOCK CENTER NEWS *The Bloomington Stock Center will be closed the week of September 13, 1993. Requests received by noon CDT on September 9 will be shipped as usual on the 13th. Orders placed between noon on the 9th and noon on the 23rd will be shipped on September 27. Please plan accordingly. *Second chromosome lethal P-inserted stocks from the Drosophila Genome Center will soon be available. Third chromosome inserts should be available sometime this fall. Less well characterized insertion strains or those containing less versatile constructs will be discarded at some point in favor of the new stocks. Individuals who contributed the stocks will be notified personally before stocks are discarded and a list of all stocks to be discarded will be posted on the BIOSCI/Bionet Drosophila newsgroup (see DIN Vol. 10) six weeks before stocks are to be discarded. Discard lists will also be posted on FlyBase in the file stock-center- news.doc in the directory flybase/stocks/stock-centers/bloomington. If you would like to receive a copy of stocks on the discard list please order them as soon as possible after the list is made public. *HAVE YOU MOVED? If you have a new mailing address please EXPLICITLY state this when you order stocks, or let Kathy know as soon as you move so our files can be updated. Address labels are generated by computer from our master address file and are not checked individually against each order. Flies sent to the wrong address create delays for you and extra work for us so please help us keep our records current. *** REQUESTS FOR MATERIALS INSERTS OR LETHALS IN 17AB Lee Fradkin, Nusse Lab, HHMI, CMGM B269, Stanford Medical Center, Stanford, CA 94305. FRADKIN@CMGM.STANFORD.EDU. We would appreciate strains or information about strains with enhancer traps or lethals that map to the 17AB region. Thanks. *** DEFECIENCY Rob Jackson, Worcester Foundation for Experimental Biology, 222 Maple Ave., Shrewsbury, MA 01545. JACKSON@SCI.WFEB.EDU. I'm looking for Df(3L)Mg27 produced by Mglinetz. I would appreciate hearing from anyone who has this deficiency or knows of its whereabouts. *** MALE STERILES Peter Clyne, Dept. of Biology, Yale U., PO Box 6666, New Haven CT 06511, USA. 203-432-3542, Fax/5631, DRONGO@VENUS.CIS.YALE.EDU I am seeking ms(2)E8 and ms(2)E9 flies which were first isolated by Edmonson in 1951. Both flies and information about whom to contact directly for the flies would be deeply appreciated. Thank you. *** MATERIAL FOR FLY THESAURUS Joanne Martinez, U. of Arizona School of Library Science/Dept. Management Information Systems. JPMARTIN@CCIT.ARIZONA.EDU Our group is developing an automatic thesaurus for the Drosophila research community. We are gathering "object filters" (gene names, protein function names, researcher names, technique names, subjects and other keywords), as well as full text electronic documents for cluster analysis (semantic relationships). The flybase and online Redbook have been very helpful for the former, but we need more electronic full text documents for determining relationships between words. I ask your assistance in providing us with a wide range of full text documents in *electronic* format, particularly abstracts and review-type articles. This is important, so that the terms in our thesaurus are as rich as possible, i.e. that the thesaurus covers as much of the terminology used in Drosophila research as possible. The documents will *not* be used for their intellectual content, so there is no problem with copyright. We are only interested in the terms and their relationships to one another. The documents will be erased once we have extracted the objects and conducted the cluster analysis. Please send whatever files you can to: jpmartin@ccit.arizona.edu Thank you very much. *** BACKISSUES OF DIS Kathy Matthews, Dept. of Biology, Indiana U., Bloomington, IN 47401. 812-855-5782, FAX/2577, MATTHEWK@INDIANA.EDU The Bloomington Stock Center would like to obtain a complete set of Drosophila Information Service. We have all volumes from number 34 to the present. If you have any of the earlier issues that you are ready to part with we would be very happy to give them a good home. *** MATERIALS AVAILABLE CHROMOSOME III HOMOZYGOUS BY DESCENT LINES Ananias Escalante and Francisco Ayala, Dept. Ecology and Evolutionary Biology, U. of California, Irvine, CA 92717. FAYALA@ORION.OAC.UCI.EDU, AESCALAN@DARWIN.BIO.UCI.EDU We are currently developing around 400 lines of D. melanogaster homozygous by decent at the chromosome III using the lethal balanced TM3 strain. These flies were collected in northern California as part of a project directed to study polymorphism in natural populations. The lines were started with males collected directly from the field. If any person is interested in these lines please contact us. These lines will be discarded after we finish our project. *** TECHNICAL NOTES DOUBLE-SIDED STICKY TAPE FOR EMBRYO INJECTIONS Mary Whiteley and Judith A. Kassis, Food and Drug Administration, 8800 Rockville Pike, Bethesda, Maryland, 20892. 301-496-9309, FAX /4684, KASSIS@HELIX.NIH.GOV One of the most common problems associated with microinjection of Drosophila embryos is the toxicity of different batches of double-sided sticky tape. Recently, we have found that batches of 3M double-sided tape purchased at our local grocery stores is often toxic to embryos. This is evident the morning after microinjection with the embryos being noticeably caved in and hollow. We called the 3M company to see if they manufactured alternate kinds of double-sided tape, and the reply was yes.... some 200 kinds. We then spoke with someone in the 3M testing laboratory about which of these 200 kinds of tape was likely to be the least toxic. She sent us a sample of industrial double- sided stick tape that had been rigorously tested for toxicity (to what we are not sure). This tape is an acetate based tape (type 415, 1/4"). We obtained a trial sample of the tape and, in our hands, and others on the NIH campus, this tape is virtually non- toxic to Drosophila embryos - we now routinely obtain about five- fold more hatching larvae than before. We have not rigorously tested this tape versus the tape at the grocery stores, nor have we tried different batches of the tape, but we think that it is important to inform Drosophilists of our success so that others may hopefully benefit. Unfortunately, 3M does not supply this tape directly, so you must first call 3M to locate the distributor in your area (612-733- 1110; ask for product information). Some distributors are reluctant to sell you less than 1 case (144 rolls). Although I have contacted some that will split up a case, the cost of a 36 yard roll is about $8.00 versus $3.00 a roll bought by the case. *** AVAILABILITY OF MATERIALS FROM THE DROSOPHILA GENOME CENTER Gerald M. Rubin, Dept. of Molecular and Cellular Biology, Life Science Annex Bldg., Box 539, U. of California, Berkeley, CA 94720. 510-643-9945, FAX /9947, FLYGENOME@MAILLINK.BERKELEY.EDU The following is an excerpt from a text document now available on FlyBase, P1.doc - further information, as well as the list of P1 clones, are available on FlyBase. The overall goal of the Center, that was funded by the NIH for three years starting August 1, 1992, is to build an integrated physical, genetic, and cytogenetic map of the Drosophila genome based on STS content mapping. We hope that the data we generate as the map is gradually assembled will be useful and widely available to Drosophila workers. In return, we ask for your help in bringing any errors, inconsistencies or independent confirmations of the data contained in these tables to our attention. Such feedback will improve the quality of the final map and speed its completion. Correspondence can be sent by email to flygenome@maillink.berkeley.edu or by FAX to 510-643-9947; alternatively correspondence can be directed to specific members of the Center as outlined below. The generation of the data described in this database is supported by a Drosophila Genome Center Grant from the NIH (NIH grant HG00750, Principal Investigator Gerald M. Rubin; Co- Investigators: Daniel Hartl, Christopher Martin, Michael Palazzolo, and Allan Spradling), by the Howard Hughes Medical Institute through its support of the Rubin and Spradling laboratories and by the DOE through its support of the LBL Human Genome Center, the home of the Palazzolo and Martin laboratories. Please acknowledge the Drosophila Genome Center in publications using this information; additional acknowledgments that apply to specific subsets of the data are detailed below. In addition to these data tables two types of materials are being made available to the Drosophila community: Clones of D. melanogaster DNA in the P1 vector and fly stocks carrying mapped single P element insertions that inactivate a vital gene. All clones, fly stocks and other information may be used for research purposes without restriction. The average insert size in the P1 library is about 80kb. The following 16 laboratories have volunteered to store the 9,216 arrayed clones that comprise the basic P1 library (approximately 5-hit) and to make them available to other laboratories in their geographical areas. They are not being financially compensated for their effort and we all owe them our appreciation. Sean Carroll, University of Wisconsin-Madison, Howard Hughes Medical Institute, Laboratory of Molecular Biology, 1525 Linden Drive, Madison, WI 53706, Tel: 608-262-3203, Fax: 608-262-4570 Allan Spradling, Carnegie Institute Washington, Howard Hughes Medical Institute, Department of Embryology, 115 West University Parkway, Baltimore, MD 21210, Tel: 410-554-1221, Fax: 410-243-6311 Hugo Bellen,Baylor College of Medicine, Howard Hughes Medical Institute, One Baylor Plaza, Room T634, Houston, TX 77030, Tel: 713- 798-5272, Fax: 713-797-6718 Steven Henikoff, Fred Hutchinson Cancer Research Center, Howard Hughes Medical Institute, Department of Genetics, Room A1-111, 1100 Fairview Avenue North, Seattle, WA 98109, Tel: 206-667-4514, Fax: 206-667-5889 Thomas C. Kaufman, Indiana University, Howard Hughes Medical Institute, Department of Biology, Jordan Hall, Room A-507, Third Street and Faculty, Bloomington, IN 47405, Tel: 812-855-3033, Fax: 812-855- 2577 S. Larry Zipursky, University of California at Los Angeles, HHMI/5-748 MacDonald Bldg., 10833 Le Conte Avenue, Los Angeles, CA 90024- 1662, Tel: 310-825-2834, Fax: 310-206-3800 Carl Thummel, University of Utah, Howard Hughes Medical Institute, Department of Human Genetics, Eccles Institute of Human Genetics Bldg. 533, Room 2100, Salt Lake City, UT 84112, Tel: 801-581-2612, Fax: 801-581-5374 Tsuneyuki Yamazaki, Department of Biology, Faculty of Science, Kyushu University, Fukuoka 812, Japan, Tel: (81)-92-641-1101, Fax: (81)-92- 632-2741 Michael Ashburner, Department of Genetics, Cambridge University, Downing Street, Cambridge CB2 3EH, England, Tel: (44)-223-333969, Fax: (44)-223-333992 Spyros Artavanis-Tsakonas, Yale University, Howard Hughes Medical Institute, Boyer Center for Molecular Medicine, 295 Congress Avenue, New Haven, CT 06536-0812, Tel: 203-737-4466, Fax: 203-787-3364 Gary H. Karpen, Salk Institute, Department of MBVL, PO Box 85800, San Diego, CA 92186-5800, Tel: 619- 453-4100, X473, Fax: 619-457- 4765 Gerald M. Rubin, University of California at Berkeley, Howard Hughes Medical Institute, Department of Molecular & Cell Biology, Room 539 LSA Bldg., Berkeley, CA 94720, Tel: 510-643-9945, Fax: 510-643-9947 Daniel L. Hartl, Harvard University, Department of Organismic & Evolutionary Biology, 16 Divinity Avenue, Cambridge, MA 02138, Tel: 617-496-3917, Fax: 617-496-5540 Matthew Scott, Stanford University School of Medicine, Department of Developmental Biology, Beckman Center, B300, Stanford, CA 94305- 5427, Tel: 415-725-7680, Fax: 415-723-9878 Paul Lasko, McGill University, Department of Biology, 1205 Ave Docteru Penfield, Montreal, PQ H3A 1B1, Canada, Tel: 514-398-6721, Fax: 514-398-5069 Marek Mlodzik, European Molecular Biology Laboratory, Differentiation Programme, Meyerhofstr. 1, D-6900 Heidelberg, Germany, Tel: (49)-62- 21-387-303, Fax: (49)-62-21-387-306 Drosophila stocks carrying mapped single P element lethal insertions are being deposited in the Bloomington Drosophila Stock Center. The first 500 lines will be available this summer or fall. More details on the P element lines will appear on FlyBase when the stocks are available. CONSTRUCTION AND REPLICATION OF THE P1 LIBRARIES. The two P1 libraries we are using were constructed by David Smoller in the Hartl laboratory. Source material for the P1 libraries consisted of Sau3A partial digests of adult genomic DNA from a highly inbred strain of genotype: y; cn bw sp. In order to minimize ambiguities in the PCR assays resulting from duplicate clones, the master library, of which copies are being distributed, was derived from the original libraries by streaking each of the 9216 clones for single colonies and then repicking individual colonies. The restreaking and library replication was carried out at LBL. RETRIEVAL OF P1 CLONES FROM THE LIBRARY. A common question about the library is about nomenclature. There are 96 microtiter plates in the library. In each microtiter plate there are 96 wells and they are labeled in an alphanumerical fashion. However, in our clone lists we describe the positions using two numbers. The first number in our clone identification refers to the microtiter plate. The second number is a conversion number for the alphanumerical position in the plates. The conversion system that we use can be most easily described as reading a book. Counting begins with the well in the upper left hand of the plate. Thus, position A1 becomes 1. The numbers increase as you move from left to right in the first row. A2 becomes 2, A3 is 3 etc. until you reach A12, which is 12. In the second row we begin counting at the left hand well at 13, thus B1 is 13. This continues across the row until B 12 is 24. This system continues down the plate with C1 as 25, etc. Ultimately, the bottom well on the right hand side H12 is 96. Using this system, a clone labeled 12-23 would be on the 12th plate in well B11. Another common question concerns how to isolate the clones. All the plates have been covered with a plastic plate sealer. This plate sealer should stay on the plates at all times and the plates themselves should never be thawed. If you want to isolate a clone, it is possible to pierce the plate sealer over an individual well with a sterile needle, or a sterile eppendorf tip. The clones can then be streaked on LB kanamycin (50 micrograms/ml of kanamycin) and grown overnight. The library was constructed in two different vectors. In each plate the clones in the wells 1 through 40 (A1 through D4) contain inserts in the P1 vector NS582 tet14ad10. In each plate the wells 41-96 (D5 through H12) contain inserts in the P1 vector ad10sacBII. References: a. General reference for the P1 cloning system: N. Sternberg, 1990 Bacteriophage P1 cloning system for the isolation, amplification, and recovery of DNA fragments as large as 100 kilobase pairs. Proc. Natl. Acad. Sci. USA 87: 103-107. b. Construction of the Drosophila NS582 tet14 ad10 library and associated methods: D. A. Smoller, D. Petrov and D. L. Hartl, 1991 Characterization of bacteriophage P1 library containing inserts of Drosophila DNA of 75-100 kilobase pairs. Chromosoma 100: 487-494. c. Description and use of the ad10 sacBII vector: J. C. Pierce, B. Sauer, and N. Sternberg, 1992 A positive selection vector for cloning high molecular weight DNA by the bacteriophage P1 system: Improved cloning efficacy. Proc. Natl. Acad. Sci. USA 89: 2056-2060. Lozovskaya, E. R., D. A. Petrov and D. L. Hartl, 1993 A combined molecular and cytogenetic approach to genome evolution in Drosophila using large-fragment DNA cloning. Chromosoma 102:253-266. d. Amplification of clone ends in ad10 sacBII by PCR: Nurminsky, D. I. and D. L. Hartl, 1993 Rapid and efficient amplification of the ends of DNA fragments cloned in bacteriophage P1. Biotechniques (in press). It is also worth noting that direct sequencing of the ends works in about 80% of the cases. Within the Center, we have been using standard cycle sequencing protocols for the ABI (dye terminator) or Pharmacia (labeled primer) automated fluorescent sequencers. For the ad10 sacBII vector we have been using the following primers: SP6L (23 bases) 5' GGCCGTCGACATTTAGGTGACAC 3'; T7L (24 bases) 5' CCGCTAATACGACTCACTATAGGG 3'. A template preparation protocol for direct radioactive sequencing of P1 ends is available from David Smoller, Genome Systems, Inc., 7166 Manchester Road, St. Louis, Missouri, 63143. It is also worth mentioning that David Smoller's company, for a reasonable price, is able to screen the same Drosophila library, as well as an additional 4000 clones with any single copy Drosophila probe. IN SITU LOCALIZATIONS OF P1 CLONES. A list of P1 clones localized to the euchromatin of Drosophila melanogaster by in situ hybridization with the salivary gland chromosomes (Oregon R) is provided. The extent of the hybridization signal is indicated by the `starting band' and `ending band' table entries. The cytology was carried out by Elena R. Lozovskaya and Robert W. Jones of the Hartl laboratory. The experimental procedures and cytological localizations have been carried out as carefully as possible, and we are confident that a very high percentage of the assignments are correct. However, in a project of this magnitude, there is an inevitable chance of error being introduced at any of a number of stages. There are also differences in judgment, for example, in deciding whether a clone with multiple sites of hybridization has one or more major sites of hybridization. In addition, there is the possibility that some mix-ups occurred in the restreaking and repicking of the library; many of the in situ hybridizations were done on the original library, prior to resteaking. There are enough cross-checks in the experimental design of the Drosophila Genome Project that any misplaced clones will be identified and corrected eventually, but users should always check for themselves. Therefore, prior to using any clone, it would be advisable to verify the cytological location by in situ hybridization. If there is any discrepancy with the assignment, please inform us immediately so that we investigate. Contact by email HARTL@MCZ.HARVARD.EDU or FAX 617-496-5854. In the event that a P1 clone you need does not match the description in the library as distributed, the Hartl laboratory can try to recover it from the original plates. In any event, we would appreciate any additional information about these clones that users could provide. A PROTOCOL FOR MAKING P1 DNA: 1. Grow an overnight culture in 25 micrograms/ml kanamycin. 2. Inoculate 500 ml of LB (25 micrograms/ml kanamycin) with 0.5 ml of the overnight culture. Shake at 325 rpm at 37 degrees C for about 3 hours (OD550 = 0.15). 3. Add 5 ml of 0.1 M IPTG (dissolve 0.6 g in 25 ml of ddH2O and filter sterilize) to the 500 ml culture. Shake for another 3 hours at 37 degrees C until OD550 = 1.3 to 1.5. 4. Harvest the cells (5K in GSA rotor for 10 minutes), and proceed with the Qiagen maxi-prep according to the kit protocol. 5. Resuspend the DNA (10-30 micrograms) in a suitable volume of TE. DATA BASED ON CONTIG ASSEMBLY. Contigs of overlapping P1 clones are being assembled by STS content mapping. The principle of physical map construction based on STS content mapping is straightforward, as shown by the following example. Consider three P1-clones denoted A, B, and C that are close together in the genome, and suppose that their content of 7 STS markers is as follows: A contains STS markers 1, 2, and 3; B contains markers 4, 6, and 7; and C contains markers 2, 4, and 5. Then it is clear that the clones must overlap, and the unique ordering consistent with the data is A-C-B (or the reverse). The STS sites are ordered within the clones as (1 3) 2 5 4 (6 7), where the parentheses around any markers indicate incomplete specification of the order. In addition, the STS content strategy requires that these single copy markers be both mapped and at least partially sequenced. In this way, in addition to identifying the overlaps between large cloned inserts, STS content mapping provides a mechanism for the introduction of biological content, flexibility and community access into the map as it is being constructed. STSs derived from the ends of mapped P1 clones and P element insertion sites are being positioned by the mapping group at LBL. STSs derived from the sequences of Drosophila genes that have been deposited in GenBank are being mapped in the Hartl laboratory. The P1 data table presents information on STSs that have been derived from the ends of the insert DNA in the P1 vector. These sequences have been generated using the SP6 primer site (S_STS) and T7 primer site (T_STS) that flank the Drosophila insert in the ad10 sacBII vector. The table also presents data on which STSs from other P1s, P element insertion sites, or known genes have been mapped to a particular P1 (Hit_by_STS). By searching for all entries with a given STS you can obtain the data necessary to assemble contigs. We hope to be able to represent these graphically in a later version of the database. The work at LBL is being jointly managed by Bill Kimmerly, Chris Martin, and Michael Palazzolo. Bill Kimmerly is the day to day supervisor of the research associates on the project.The research associates working on the mapping project at LBL are: Karen Stultz; Victor Stevko; Ami Richardson; Gail Shirley; and Dan Hong. Charles Yu is an undergraduate also working on the project. LETHAL P ELEMENT INSERTIONS The overall goal of this part of the project is the analysis of P element insertion sites that disrupt vital autosomal genes in order to cross- reference the physical, cytogenetic and genetic maps of the Drosophila melanogaster genome. By defining STS's within sequences adjacent to all those insertions disrupting different vital genes, this collection would serve as a versatile link between the genetic and physical maps of the Drosophila genome. There are thought to be about 4,000 autosomal Drosophila genes capable of mutating to lethality. Our original project involved a collection of 1,800 autosomal recessive lethal P insertion lines, that were expected to define approximately 1,100 vital genes on the physical map, or about 27% of the total. Our goal is to create a collection of 1,000-1,200 Drosophila strains meeting specific quality criteria. Lines should contain single P element insertions each defining a unique vital gene. A group of 1,800 candidate strains has been assembled; non- redundant, single- insert strains causing recessive lethality will be selected and mapped from among these lines through seven sequential steps: 1. Map the chromosome location of the insertion in each of the 1,800 lines by in situ hybridization to polytene chromosomes. 2. Identify and eliminate lines containing background lethal mutations from the initial collection of 1800 lethal single P insertion strains. 3. Identify and remove lines with two insertions, and also redundant lines, in which the same gene is mutated, from the collection of 1800 lethal single P insertion strains. 4. Identify and eliminate lines in which the P insertion has not caused a lethal mutation. 5. Plasmid rescue DNA flanking the P element insertion from each of the approximately 1,100 lines that are expected to remain after the criteria of specific aims 2-4 have been applied. 6. Determine the sequence of approximately 400 bp of genomic DNA immediately adjacent to the site of insertion in each of these lines to provide an STS for mapping onto the P1 library. We have collected pre-existing lethal lines from several laboratories as the starting material for this project. These initially included the Spradling, Rubin, Scott and Jan laboratories. More recently we have initiated a collaboration with Istvan Kiss which will allow us to substantially increase the number of P-induced lethals available for the project. We would appreciate hearing from individuals with collections of P- induced lethal, sterile, or visible mutations that they would like to make available. Contact Allan Spradling (email:spradling@mail1.ciwemb.edu or FAX 410-243-6311). The in situ hybridization localization of P element insertion sites (Step 1) is being carried out in the Rubin laboratory by Todd Laverty, Glenn Doughty, Wan Yu and Donna Nakahara. The Genetic verification tests (Steps 2 and 3) are being carried out in the Spradling laboratory by Allan Spradling and Dianne Stern. The first collection analyzed was that from the Spradling laboratory. Lines will be deposited in the Bloomington Stock Center when steps 1-3 above have been completed and will be posted here at that time. We expect the first 500 lines to be available from the Stock Center sometime this summer. References: The isolation of the P element lines from the Spradling laboratory was described in: Karpen, G.H. and Spradling A.C. (1992). Analysis of subtelomeric heterochromatin in the Drosophila minichromosome DP1187 by single-P-element insertional mutagenesis. Genetics 132: 737- 753. The PZ-enhancer trap element used to generate the Spradling lab lines is described in Mlodzik, M. and Hiromi, Y. (1991). The enhancer trap method in Drosophila: its application to neurobiology. In: Gene Expression in neural tissues. Methods in Neuroscience, Vol 9. P.M. C. Orlando, ed. Academic press. *** GENETIC NOTES CORRECTIONS FOR THE REDBOOK Dan Lindsley and Georgianna Zimm, Dept. of Biology, U. of California, La Jolla, CA 92093. 619-534-3109, FAX/0053, REDBOOK@JEEVES.UCSD.EDU, ZIMM@JEEVES.UCSD.EDU (p=page; L=left; R=right) p 215L, fj (cytology): "Df(2R)11B" to "Df(2R)Pcl11B" p 215L, fj (cytology): "Df(2R)Pcl-w5" to "Df(2R)Pcl-W5" p 548L, "Phb: Photophobe" to "Ppb: Photophobe" p 548L, change all table and text entries from "Phb" to "Ppb" p 549L, "Photophobe: see Phb" to "Photophobe: see Ppb" p 702L: remove the references for the gene tbs p 782L, wt (alleles): Insert this reference (after left parenthesis) "Mglinetz and Vikulova, 1977, Genetika 13: 1318-20;" p 782R, wt (cytology): Second line should be "Df(2R)Pcl-W5 = Df(2R)55A-B;55C but not Df(2R)Pcl11b" p 854L, Df(2R)Pcl (table): "Df(2R)Pcl11B[beta]" to "Df(2R)Pcl[beta gamma]" p 854L, Df(2R)Pcl (table): "Df(2R)Pcl-W5" to "Df(2R)Pcl-W5[delta]" Note: Page corrections made in Photophobe corrections of Ballinger. Editor's note: A complete list of reported corrections to the Redbook and to Ashburner's Greybook are maintained on FlyBase. See flybase/greybook/errors.txt and flybase/redbook/update.txt. A more coherent version of the latter will be available soon as redbook/errors.txt. -- K.M. ***