DROSOPHILA INFORMATION NEWSLETTER Volume 2, April 1991 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 for publication in DIS (see below). Materials appearing in the Newsletter will be reprinted, in unedited form, in the next issue of DIS. Material appearing in the Newsletter may be cited unless specifically noted otherwise. Material for publication may be submitted in any of the following formats - Macintosh Microsoft Word or MacWrite, MS-DOS WordPerfect, or text/ASCII file. Figures and photographs cannot be accepted at present. Send material, in order of preference, as E-mail (addresses below), on floppy disk, or as laserwriter or typed hard-copy (not bit-mapped). Technical notes should be sent to Carl Thummel, all other material should be sent 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. 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 many of 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, WT 84112 812-855-5782; FAX/2577 801-581-2937; FAX/5374 MATTHEWK@IUBACS.BITNET THUMMEL@MEDSCHOOL.MED.UTAH.EDU MATTHEWK@UCS.INDIANA.EDU *** To add your name to the Newsletter distribution list, send one of the following E-mail messages. 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 are on the list and do not wish to receive DIS, or you want to remove a 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. 2 TABLE OF CONTENTS >Introduction to Drosophila Information Newsletter >How to subscribe to the Newsletter >TABLE OF CONTENTS >ANNOUNCEMENTS >Falkenthal Memorial Fund >DIS Vol. 69 - The Genetic Maps of Drosophila >DIS Vol. 70 - Traditional DIS >33rd and 34th Drosophila Research Conferences >1991/1992 Drosophila Board >Wisconsin Drosophila Genome Meeting Summary Report >REQUESTS FOR MATERIALS >Clones for Drosophila genome project >Survival data >Deficiencies for the Bloomington Stock Center Subject: Correction to DIN Vol. 2 The request for material "Deficiencies for the Bloomington Stock Center" appearing in DIN Vol. 2 contains a typo. The gap in the deficiency collection shown as 44E2-44C3 should read 44E2- 46C3. >DATABASES/COMPUTING >IUBIO - Drosophila information from the network >GENETIC NOTES >A modest proposal - Nomenclature for transcription factors >Autosynaptics made easy >TECHNICAL NOTES >New pCaSpeR P element vectors >Cloning with YACs >Effect of growth medium on ADH major/ADH minor activity *** ANNOUNCEMENTS FALKENTHAL MEMORIAL FUND Our colleague Scott Falkenthal died last fall. It was Scott's wish that donations in his memory should be put into an endowment fund that will be used by the Department of Molecular Genetics at Ohio State University to support their annual Graduate Student Colloquium and departmental seminar program. Donations can be sent to: Scott Falkenthal Memorial Fund, c/o Dr. Lee Johnson, Dept. of Molecular Genetics, Ohio State Univ., 484 West Twelfth Ave., Columbus, OH 43210-1292. *** DROSOPHILA INFORMATION SERVICE Vol. 69 William Gelbart, Biological Labs., Harvard Univ., 16 Divinity Ave., Cambridge, MA 02138-2097, USA. 617-495-2906, FAX/9300. The Genetic Maps of Drosophila, compiled by Michael Ashburner, will be published in mid-May 1991 as DIS 69. The issue is a 400 page compendium of five tables representing the genetic maps of Drosophila. GENETIC LOCI is a list of the genes sorted alphabetically. GENETIC MAP is a genetic map sorted by genetic map position. GENETIC FUNCTION is a list of loci sorted by the "function" of the gene's product. SYNONYMS is a list of synonyms with their "valid" genetic symbols. REFERENCES is a table of references to these data, usually updating Lindsley and Zimm. Copies of DIS 69 must be prepaid by check in U.S. currency drawn on a U.S. bank. Make checks payable to DIS-69. Purchase orders or standing orders cannot be accepted. Domestic shipment will be by United Parcel Service (UPS) and delivery should take about one week from time of shipment. Foreign orders may be placed for delivery by surface mail or air parcel post. Surface mail will take 6-8 weeks for delivery, while air parcel post will take 1-2 weeks, depending upon destination. Note however that foreign air mail will significantly increase the purchase price. Purchase requests should be received by April 15, 1991. The size of the print run will be determined at that time by the number of copies ordered at that time. We anticipate that the issue will be mailed by the end of May. Price per copy is $13.00 plus shipping. Add shipping costs as follows: Domestic UPS - $3.00 per copy. Foreign surface mail - $6.00 per copy. Foreign air parcel post to: Canada - $7.00 per copy; Mexico and Central America - $13.00 per copy; South America - $21.00 per copy; Europe - $23.00 per copy; other foreign destinations - $27.00 per copy. Submit purchase requests or make inquiries to DIS-69, c/o William Gelbart, Editor, at the address above. *** DROSOPHILA INFORMATION SERVICE Vol. 70 James Thompson, Dept. of Zoology, Univ. of Oklahoma, Norman, OK 73019. 405-325-4821; FAX/7560. Drosophila Information Service has a long and respected history of promoting communication among geneticists, ecologists, systematists, and molecular biologists who focus upon this species in their research. DIS is a non-profit service to biologists world-wide. As you know, Philip Hedrick has stepped down after many years as editor of DIS. I hope you will join me in thanking him for his efforts. In the interests of maintaining this useful organ of communication for the Drosophila community, I have volunteered to become the next editor of DIS. The format of DIS will remain very similar to those in the past, but some areas will be added or expanded. * Research notes and new mutant sections will continue. * Coverage of techniques will be expanded to include more molecular and cellular techniques. * A group of assistant editors will be invited to help solicit material and develop new areas of coverage. * International representatives will be included to help draw attention to Drosophila activities world-wide. * Information will be provided on accessing and using new sources of information, such as the Drosophila Information Newsletter and computerized stock and clone lists. * Information on Drosophila Research conferences in the U.S. and abroad will be included to help promote better communication among all Drosophila workers. DIS 70 may be ordered by sending a check for $8.00 (U.S. dollars drawn on a U.S. bank, payable to "Drosophila Information Service") per copy to the above address. *** 33RD and 34TH DROSOPHILA RESEARCH CONFERENCES The 1992 Drosophila Research Conference will be held March 10-15th at the Wyndham Franklin Plaza Hotel in Philadelphia, PA, USA. Bill Gelbart will chair the organizing committee. The 1993 meeting was scheduled to meet at Asilomar, but the large turn out for Chicago (over a 1000 registered participants; Asilomar has a theoretical capacity of 800) has caused the board to reconsider. A final decision has not yet been made, but the choice is between Asilomar and San Diego. Dates will be announced when available. Gerry Rubin will be the organizing committee chair for the 1993 meeting. *** YOUR REPRESENTATIVES ON THE GSA DROSOPHILA BOARD The Drosophila Board of the GSA meets every year (and communicates more often by phone) to decide on issues of interest to the fly community. These issues center, for the most part, around the arrangements for the annual fly meeting organized by the GSA, but also cover a variety of other topics, such as the scope and format of DIS. If you have any questions or suggestions for the Board, please feel free to contact your local representative, listed below. Maine, Vermont, New Hampshire, Massachusetts, Connecticut, Rhode Island: Terry L. Orr-Weaver, Whitehead Institute, Biomedical Research, 9 Cambridge Center, Cambridge, MA 02142. (617) 258-5245, (617) 258-5061 FAX Downstate New York, New Jersey, Eastern Pennsylvania, Delaware, West Virginia, Washington, DC, Maryland, Virginia: Claire Cronmiller, Dept. of Biology, Gilmer Hall Rm. 229, Univ. of Virginia, Charlotttesville, VA 22901. (804) 924-7937, (804) 982-2653 FAX North Carolina, South Carolina, Georgia, Florida, Alabama, Mississippi, Kentucky, Tennessee, Louisiana, Puerto Rico: John C. Lucchesi, Dept. of Biology, Univ. of North Carolina, Chapel Hill, NC 27599. (919) 962-1332, (919) 962-1625 FAX Minnesota, Wisconsin, Iowa, Illinois, Indiana, Missouri: Kathleen Matthews, Dept. of Biology, Indiana University, Bloomington, IN 47405. (812) 855-5782, (812) 855-2577 FAX Upstate New York, Toronto, Ohio, Western Pennsylvania, Michigan: Mariana F. Wolfner, Section of Genetics and Development, Biotechnology Building, Cornell University, Ithaca, NY 14853. (607) 254-4801, (607) 255-2428 FAX Utah, Colorado, Kansas, Nebraska, North Dakota, South Dakota, New Mexico, Texas, Arizona, Oklahoma, Arkansas, Louisiana: Steve Cohen, Dept. of Cell Biology, Baylor College, 1 Baylor Plaza, Houston, TX 77030. (713) 798-5019, (713) 790-1275 FAX Oregon, Washington, Idaho, Montana, Wyoming, Alaska: Steven Henikoff, Hutchinson Cancer Research Center, 1124 Columbia St., Seattle, WA 98104. (206) 667-4515 California, Hawaii, Nevada: John R. Merriam, Dept. of Biology, University of California, Los Angeles, CA 90025. (213) 825-2256, (213) 206-3987 FAX Canada: Thomas Grigliatti, Department of Zoology, Univ, of BC, #2354-6270 University Blvd., Vancouver, BC, Canada, V6T 2A9. (604) 228-2161 *** DROSOPHILA GENOME MEETING SUMMARY REPORT - ABSTRACT (the complete report is posted on IUBIO in ARCHIVE.FLY.NEWS) William Reznikoff, Dept. of Biochemistry, Univ. of Wisconsin, Madison, WI 53706-1569, USA. 608-262-3026. Drosophila melanogaster is one of the prime model organisms for the human genome initiative. In order to develop an outline of the current state of genome analysis in the Drosophila community, to suggest some possible goals of Drosophila genome research and to consider possible strategies for achieving these goals, a workshop was held in Madison, Wisconsin, on August 3-5, 1990, with the support of the National Center for Human Genome Research, NIH, and the University of Wisconsin Graduate School. Although the participants in the workshop were limited in number, the organizers did try to ensure that scientists from various specialties and a variety of institutions were invited to achieve a reasonable representation of the Drosophila research community. In addition to scientists from the Drosophila community, the workshop was also attended by representatives from the C. elegans genome initiative, and by experts in sequence technology and biocomputing. The workshop participants decided to publish a summary version of its report because of the importance of the issues raised and the limited participation. We invite comments. The workshop started with a challenge--why initiate a large scale genome project during a time of tight funding for very productive individual projects? The workshop participants returned to this concern throughout the meeting. The workshop participants felt that if any funds were being allocated for the genome initiative, then the Drosophila projects should represent a high priority for those funds. Drosophila is unique amongst multicellular organisms for the genetic information available and genetic, molecular and developmental research currently being pursued. This means that well designed sequencing projects targeted on any of several genetically well-characterized chromosome regions are likely to yield important data which are easily interpreted and of significant usefulness to the research community. Many of the complex biological processes found in humans are also found in Drosophila. Its powerful genetic system coupled with a sequence analysis of selected regions is likely to provide important clues to the genetic control of the analogous processes in humans. Drosophila is also unique in that the polytene chromosomes already provide a high resolution physical map to which the molecular map may be easily aligned. The P element and other similar transposons are already proven to be powerful tools for further enhancing genetic research and provide new methods for accessing the DNA for physical mapping and sequence analysis. This technology can serve as a model for similar approaches in other organisms. Finally, the Drosophila genome is of a size which allows one to reasonably consider a complete sequence analysis. The workshop included reports from currently funded and proposed Drosophila genome projects, discussion of genome analysis work being done on the C. elegans and E. coli systems, reports on the current initiatives to manage information and strain maintenance and distribution, and a description of advanced sequencing technologies. The workshop participants wrote a report which is available through the National Center for Human Genome Research, NIH. The fruits of science must be increased public knowledge. The genome initiative presents unusual challenges in realizing this obligation. Ongoing genome projects should be configured such that they share information with each other and make available in a timely fashion data to the Drosophila community. This means that a high priority should be placed on the development and maintenance of informatics systems which can provide many forms of access, including hard copy and on-line reports. It is hoped that a frequently updated database relating known genes, P element insertion points, genetic rearrangements, polytene map locations, YAC and cosmid maps, and sequences, will be available. In addition, the participants in this workshop found that this type of meeting could also serve an important role in sharing the genome project with the Drosophila community. We suggest that annual workshops should be part of the genome initiative. It is important that such meetings have attendance by members of the Drosophila community who are not directly involved in genome projects. Such participation will help disseminate the fruits of the initiative and will help to refine the priorities to reflect the changing needs of the community. *** REQUESTS FOR MATERIALS REQUEST FOR CLONES Leonard Rabinow[1] and Robert Saunders[2]. [1]Waksman Institute, Rutgers Univ., Piscataway, NJ 08855-0759, USA. 908-932-0091/0092, FAX/5735, RABINOW@BIOVAX. [2]Dept. of Biochemistry, Univ. of Dundee, Dundee DD1 4HN, UK. (0382)23181 ext. 4790, FAX/201063, BI31@UK.AC.DUNDEE.PRIMEB (reverse node order from USA). Most readers of DIS are probably aware of a number of projects in progress with the aim of constructing a physical map of the Drosophila melanogaster genome. A collaboration among the laboratories of F.C. Kafatos, C. Louis, C. Savakis, D.M. Glover, and M. Ashburner is proceeding by fingerprinting cosmids selected with cytological division-specific probes produced by microdissection from polytene chromosomes (Nucl. Acids Res. (1990) 18, 6261-6270). J. Messing, L. Rabinow, W. Sofer and G. Hamm at the Waksman Institute, are initiating a project to produce a restriction map of the genome, using bacteriophage P1 and automated reading of partial restriction digests. Both projects will correlate their developing physical maps with the genetic and cytological maps of Drosophila. To achieve this, we would like to probe our libraries with loci cloned and mapped by members of the Drosophila community. We would greatly appreciate it if members of the fly community willing to contribute clones would send them to either of the above addresses. It would be most convenient if clones could be sent in plasmids (preferably bearing T7, SP6 or T3 RNA polymerase promoters), or M13, to avoid cross-hybridization problems. A few micrograms of a specific oligonucleotide, if available, could also be used. Submissions would be most useful if accompanied by information on the name, identity, and location of the clone, its vector, a restriction map or sequence, and a reference. To coordinate the overall efforts, clones and data will be exchanged among these groups. Submitted clones will be exchanged only between these two groups, unless specifically authorized by the sender. *** DEFICIENCIES FOR THE BLOOMINGTON STOCK CENTER Kathy Matthews, Dept. of Biology, Indiana Univ., Bloomington, IN 47405. 812-855-5782, FAX/2577, MATTHEWK@IUBACS. The Drosophila collection at the Bloomington Stock Center currently carries 512 deficiency stocks. Our "Deficiency Kits" consist of the minimum set of these deficiencies that cover the maximum amount of the genome. The kits for the X and the three autosomes currently consist of 142 stocks - 40 in DK1, 49 in DK2, 51 in DK3, and 2 in DK4. Together these lines provide coverage for about 60% of the euchromatin. We would like to fill as many of the existing gaps in coverage as possible. Regions of the polytene chromosomes that are either NOT covered by the current collection, or the breakpoint uncertainty for deficiencies we do have is such that coverage is doubtful, are shown below. If you have deficiencies that help fill any of these gaps and are willing to share them with your colleagues, please send them to the stock center. Subject: Correction to DIN Vol. 2 The request for material "Deficiencies for the Bloomington Stock Center" appearing in DIN Vol. 2 contains a typo. The gap in the deficiency collection shown as 44E2-44C3 should read 44E2- 46C3. 005A01-005A08, 005D05-005E03, 006B01-006E04, 007C03-007D01 007D05-007D10, 008C06-008E12, 011F01-011F10, 012A01-012A10 012F05-013F01, 014B01-014B06, 015A06-017A01, 018A02-018A05 021B07-021C01, 021D05-024A03, 025C08-025D02, 026A08-027E08 028C01-031A03, 032C01-032F01, 034A01-034B12, 035E06-036A08 036E01-036E04, 040A04-041F11, 041A01-042A16, 044E02-044C03 046C09-047D03, 049F15-051A08, 051B06-051C03, 052F10-055A01 055F01-056D15, 056F01-056F09, 058B01-059D08, 060A07-060C06 060D09-060E03, 061A01-062B07, 062B12-062F12, 063E01-066D10 066E01-066F05, 067D07-067F03, 069B04-070A03, 070A05-070B07 070D06-070D07, 074F01-075B03, 075C01-076A03, 076B02-077A01 077F01-078A03, 079A04-080E03, 081F01-083C01, 085B06-085D08 085F06-086C01, 088E05-089A01, 090A01-090D01, 092D03-093B05 094A01-094F06, 095A01-096A07, 096A21-097A10, 098A01-099B11 100B04-100F05, 101A01-101E09, 102B10-102E02, 102E10-102F17 *** DATABASES/COMPUTING IUBIO - DROSOPHILA INFORMATION FROM THE NETWORK Kathy Matthews, Dept. of Biology, Indiana Univ., Bloomington, IN 47405. 812-855-5782, FAX/2577, MATTHEWK@IUBACS. A variety of information useful to Drosophilists is posted on IUBIO, our Department of Biology micro-VAX, and is available to the public via anonymous FTP (File Transfer Protocol) on the Internet. In addition to current stock lists from the Bloomington Stock Center, the following Drosophila-oriented information is maintained on IUBIO: Michael Ashburner's genetic maps (if you don't want to wait for DIS 69, or need an update later) and his most recent Drosophila codon bias table; John Merriam's GenMaps database (generally known as the clone list, but contains a lot of other useful information in addition); news items such as back issues of DIN and the full text of the report from the Wisconsin genome meeting. If you are interested in posting information on IUBIO, contact IUBIO founder and archivist, Don Gilbert. Don can be reached at ARCHIVE@IUBIO.BIO.INDIANA.EDU (preferred address) or GILBERTD@IUBACS. To retrieve information from IUBIO, the following instructions work for most people/computers, but note that you must have access to the Internet. From your system prompt, type: FTP IUBIO.BIO.INDIANA.EDU (this connects you to IUBIO using FTP software on your local computer) user: ANONYMOUS (spell it correctly or it won't work) password: YOUR REAL USER NAME (anything works) CD [.FLY] (cd = change directory; Drosophila info is in the FLY directory) DIR (this shows file and subdirectory names) GET FILENAME (use this command to copy one file at a time) or MGET *.README (to get all of the readme files; you will be asked to confirm each file, but it's still easier than typing each filename; MGET doesn't work between the VAX and a PC) MGET *.TXT (to get all of the data files) BYE (to log off) Ashburner's map lists are in the subdirectory LOCI. To copy these, type CD [.FLY.LOCI] and continue. His data files have the extension .TEXT instead of .TXT, so use the command MGET *.TEXT for these files. For news items, type CD [.FLY.NEWS] and use GET FILENAME to copy the files you are interested in. If you want files placed in a particular subdirectory on your local account, type LCD SUBDIRECTORY NAME (the complete path if you are working from a PC) before giving the GET command. *** GENETIC NOTES A MODEST PROPOSAL - NOMENCLATURE FOR TRANSCRIPTION FACTORS Dan Lindsley, Dept. of Biology, Univ. of California, La Jolla, CA 92093. 619-534-3109, FAX/0053. A modest proposal: As biochemically identified transcription factors in Drosophila are cloned and mapped by in situ hybridization to polytene chromosomes they be designated by Trf followed by the polytene location of the encoding gene. This nomenclature would apply to gene products whether they are subunits of heteromultimeric proteins or function as monomers or homomultimers. Thus, for example, the gene named Elf-1 by Bray, Burke, Brown, and Hirsh (1989, Genes Dev. 3:1130-45) and Ntf by Dynlacht, Attardi, Admon, Freeman, and Tjian (1989, Genes Dev. 3:1677-88), and so designated in the upcoming revision of the Red Book, would be designated Trf54F. Major developmental mutants shown to function in the regulation of transcription would remain named according to the phenotype of the mutants by which they were originally identified. *** AUTOSYNAPTICS MADE EASY David Gubb[1], John Roote[1], Darin Coulson[1], Claire Henchcliffe[2], Terry Lyttle[3] and Bruce Reed [1]. [1]Dept. of Genetics, Univ. of Cambridge, Cambridge CB2 3EH, UK. 44-223- 333969, FAX/333992, DG27@UK.AC.CAM.PHX (invert node order from USA). [2]Howard Hughes Medical Institute, Dept of Biochemistry, Univ. of California, Berkeley, CA 94720, USA. 415-642-9402, FAX/3-5548. [3]Dept. of Genetics and Molecular Biology, Univ. of Hawaii, Honolulu, Hawaii 96833, USA. 808-948-7860, LYTTLE@HUCCVX. The possibility of using autosynaptic chromosomes is a relatively new addition to the techniques available to the Drosophila geneticist. Developed by Loring Craymer through his work on the manipulation of pericentric inversions (1981, Genetics 99: 75-97), autosynaptics are attractive as an alternative to T(Y;A)s for generating synthetic duplications and deletions (Lindsley and Sandler et al. 1972, Genetics 71:157-184), but with the advantage that the heterochromatic breakpoints are autosomal. As defined by Craymer (1981), autosynaptic chromosomes are the aneuploid products of recombination between a pericentric inversion and a cytologically wild-type chromosome. On Craymer`s terminology, the left- and right-hand breakpoints of a standard "heterosynaptic" inversion are separated onto homologous centromeres to give reciprocal "levosynaptic (LS)" and "dextrosynaptic (DS)" elements. The LS element will be homozygous for the left arm chromosomal regions distal to the inversion breakpoint which will pair "autosynaptically". The complementary LS + DS elements form a euploid autosynaptic "constellation" which is indistinguishable cytologically from the same inversion in the heterosynaptic form. Among other attributes, autosynaptic chromosomes can form the basis for self-selecting screens designed to recover new chromosome breakpoints within a defined cytological region (Gubb, McGill and Ashburner, 1988, Genetics 119: 377-390). What follows is a list of available stocks that we have constructed, with suitable genetic marker mutations, and, one hopes, a simple explanation of their use. In order to simplify the construction of segmental aneuploids, the pericentric inversions used were chosen to have one breakpoint in euchromatin and the other within the centric heterochromatin. There are two advantages in this approach. Firstly, a large number of inversions of this type were available, because the centric heterochromatin is a large target area for chromosomal breakpoints. Secondly, because aneuploidy for centric heterochromatin has little effect on viability, only the position of the single euchromatic breakpoint need be considered in the construction of segmental aneuploids. When two autosynaptic stocks are crossed together there are only two potentially viable classes of progeny, one carrying a deletion and the other a duplication. Thus: LS(2)A//DS(2)A X LS(2)B//DS(2)B -> LS(2)A//DS(2)B + LS(2)B//DS(2)A. (The remaining products, LS(2)A//LS(2)B and DS(2)A//DS(2)B will be grossly aneuploid and embryonic lethal.) In our set of stocks care has been taken to ensure that these classes may be distinguished from each other by appropriate choice of markers. In many crosses only the duplication class will survive. It is, however, simple to recover additional chromosomes with breakpoints in the region surrounding an existing autosynaptic breakpoint using either P-element or X-ray mutagenesis (Gubb, McGill and Ashburner, 1988 Genetics 119:377-390). These additional stocks will allow deletion mapping of the region in question. We have listed below our set of autosynaptic stocks with euchromatic breakpoints on the left arm of chromosome two. These stocks have been sent to the Bloomington stock center from where they are available on request. The list gives the Bloomington stock designation, #, and includes one stock made by L. Craymer, # 1166. Many of the inversions used were originally isolated by W. Gelbart and co workers (thank you Bill et al.). #1166 LS(2)S[325]. In(2L)Cy, Cy E(S)//DS(2)S[325],, Sco 21F;41* #3537 LS(2)DTD18, ho[2]//DS(2)DTD18, cn bw 23A4-7;41 #2477 LS(2)DTD21, ho[2]//DS(2)DTD21, cn, {dp} 23A1.2;41 #3538 LS(2)DTD16,, dp//DS(2)DTD16, bw, dp 23B;41** # 865 LS(2)DTD8, {net}//DS(2)DTD8, sp 23C-D1;41 #3202 LS(2)DTD42, {net}//DS(2)DTD42, bw sp 23E3-6;41 #3067 LS(2)DTD52//DS(2)DTD52, vg 24D1.2;41 #2427 LS(2)DTD124,, b//DS(2)DTD124, cn, b 24D2-3;41 #3535 LS(2)DTD109, {fy[2]}//DS(2)DTD109, bw 25E2-3;41 #3536 LS(2)DTD116, {net}//DS(2)DTD116, sp 26A4-6;41 #558 LS(2)DTD24, {net}//DS(2)DTD24, bw sp 26C1.2;41A #3539 LS(2)DTD51. In(2L6)Cy//DS(2)DTD51, cn bw, {b} 27D1;41 #3540 LS(2)DTD11//DS(2)DTD11, sp 28A;41 #3541 LS(2)DTD111. In(2L)Cy, {al[2]} Cy {pr}//DS(2)DTD111, vg 29F;41AB #3551 LS(2)DTD125, net//DS(2)DTD125, bw sp 31E;41 #1660 LS(2)DTD107, In(2L)DTD107 {TE52 ho[2]}. dp//DS(2)DTD107, bw 32F;41 #3534 LS(2)DTD4, fy[2]//DS(2)DTD4, bw 32F;41AB #3552 LS(2)DTD86, Bl//DS(2)DTD86, {cn} 33B1.2;41 #3550 LS(2)b81a2//DS(2)b81a2 34D;41D//34E3;41D #2047 LS(2)D20, {net dp b}//DS(2)P9, {pk cn sp}, pr 34E4-F2;41/ /34B7-12;41D #3549 LS(2)DTD43, ho[2]//DS(2)DTD43, bw sp 35B1.2;41 #3542 LS(2)D9, dp b//DS(2)D6 35B1-3;41//35D5-7;41 #3544 LS(2)ScoR+9//DS(2)D2, cn bw; C(1)M4, y[2] 35D1-2;41/ /34D4.5;41B3-9 #2089 LS(2)D5//DS(2)CH25 36C1;42F//36C;41 #2040 LS(2)P3,,{pr}//DS(2)noc[4], cn bw 37B.2;41CD//35B1.2;41A #1052 C(2L)C3, b pr; C(2R)C1, sple 40DE;41A * S[325] breakpoint was induced within Dp(2;2)S thus both LS and DS may carry a small duplication. ** LS also carries Df(2L)DTD16 24D3-E1. {} We have used curly brackets to indicate that the enclosed allele, {net} for example, was present on one of the heterosynaptic chromosomes from which the autosynaptic chromosome was derived, and is thus heterozygous in the autosynaptic stock. The position of marker mutations relative to the inversion breakpoints is indicated by commas: LS(2)DTD125, net is homozygous for net which maps distal to the 31E breakpoint of DTD125. Similarly, DS(2)DTD124, cn, b carries cn distal to the 41 breakpoint and b proximal to the 24D2-3 breakpoint i.e., within the heterosynaptic region of the DS element. There are a few occasions, when heterozygous dominant markers are used, where this nomenclature would remain ambiguous. In such cases the position of the centromere is indicated by a period. A similar set of chromosomes for the right arm of the second chromosome is currently being constructed in Cambridge. The available stocks are: LS(2)lt[G16],, lt stw cn bw//DS(2)lt[G10],, lt cn 40;60E5-8/ /40;59F3-4 LS(2)lt[G10], b el, bw//DS(2)lt[G10],, lt {cn} 40;59F3-4 LS(2)lt[G16],, lt stw cn bw//DS(2)bw[v32g], bw, lt[v] {sp} 40;60E5-8//40;59E LS(2)bw[v32g], net bw//DS(2)bw[v32g], bw, lt[v] {sp} 40;59E LS(2)Pu[L]//DS(2)Pu[L], {or} If 40;57B-C These stocks can be obtained from Bruce Reed (e-mail: BR106@UK.AC.CAM.PHX, invert node order from USA). The Craymer stock LS(2)f6//DS(2)f6 39D3-E1;48F6-49A1 (available from Bloomington as stock #1231) fits in with this 2R series. EXAMPLES OF THE USE OF AUTOSYNAPTIC STOCKS TO GENERATE SEGMENTAL ANEUPLOIDS. 1) To generate the reciprocal duplication and deletion between 23A1.2 and 23C-D1, flies of stock #2477 are crossed to flies of stock #865. The progeny should be phenotypically cn (Dp 23A1.2;23C-D) or ho[2] sp (Df 23A1.2;23C-D). Thus: #2477 LS(2)DTD21, ho[2]//DS(2)DTD21, cn, {dp} X #865 LS(2)DTD8, {net}// DS(2)DTD8, sp --> LS(2)DTD8, {net}//DS(2)DTD21, cn, {dp} (Dp 23A1.2;23C-D) + LS(2)DTD21, ho[2]//DS(2)DTD8, sp (Df 23A1.2;23C- D). These two classes of aneuploid autosynaptic flies can be maintained as stable stocks. The corresponding heterosynaptic inversions can be recovered by mating virgin female autosynaptic flies to normal (heterosynaptic) males. Thus: LS(2)DTD8, (net)//DS(2)DTD21, cn, {dp} X CyO/Gla -> In(2LR)DTD8[L]DTD21[R], {net} dp cn/CyO (or Gla) + {net} dp cn/CyO (or Gla). It is likely, although not certain, that the net mutation will be carried on the cytologically wild-type chromosome rather than the aneuploid inversion. (The net mutation was introduced into the autosynaptic constellation by crossing In(2LR)DTD8/net sp to an autosynaptic stock. It would be possible to lose the net mutation by recombination within the LS element distal to the inversion breakpoint. This would be a rare event as the net mutation is relatively close to the breakpoint and, in practice, we have been unable to select for homozygous net flies in the autosynaptic DTD8 stock.) 2) Similarly the cross between #3534 and #3549 will give ho[2] bw flies that carry Dp 32F;35B1.2. Flies of the reciprocal class, Df 32F;35B1.2, will not survive, as deletions of this length are lethal. If deletions in the 32F;35B1.2 region were required, then the hyperploid autosynaptic stock LS(2)DTD43, ho[2]//DS(2)DTD4, bw could be used to recover additional autosynaptic elements with breakpoints within the duplicated interval, by mutagenesis (Gubb, McGill and Ashburner, 1988 Genetics 119:377-390). As in the previous example, the hyperploid autosynaptic (DTD43//DTD4) can be resolved to the equivalent hyperploid heterosynaptic inversion, In(2LR)DTD43[L]DTD4[R], by mating autosynaptic females to normal males. Unfortunately, the two resolved products of this autosynaptic are phenotypically indistinguishable, but can be identified by cytology. Hyperploid pericentric inversions such as In(2LR)DTD43[L]DTD4[R] can be particularly useful in screening for haplo-lethal or haplo- sterile mutations as they will act as balancer chromosomes for the duplicated regions. The problem of distinguishing between the euploid and aneuploid products of resolution of an autosynaptic stock without resorting to cytology does not arise with inversions that have a dominant mutation associated with one of their breakpoints. For example: w; LS(2)ScoR+1, net//DS(2)TE146-SZ4, cn, pr, which carries a w[+] rst[+] transposable element (TE146 of G. Ising) associated with the DS breakpoint. If females of this stock are crossed to w; Gla/CyO males, the euploid heterosynaptic progeny are phenotypically white-eyed while the aneuploid progeny are red- eyed. We are currently generating a set of stocks carrying the w[+] gene within a single P{w[+]} construct element at the inversion breakpoints. With these stocks, as with the above example, the w[+] phenotype will identify the aneuploid In(2LR) product of resolution of the autosynaptic stock. *** TECHNICAL NOTES NEW pCaSpeR P ELEMENT VECTORS Carl S. Thummel[1] and Vincenzo Pirrotta[2]. [1]HHMI, Eccles Institute, Bldg. 533, Univ. of Utah, Salt Lake City, UT 84112, USA. 801-581-2937, FAX/5374, THUMMEL@MEDSCHOOL.MED.UTAH.EDU; [2] Dept. of Cell Biology, Baylor College of Medicine, 1 Baylor Plaza, Houston, TX, 77030, USA. 713-798-6635, FAX/790-1275, PIRROTTA@BCM.TMC.EDU. We have modified the pCaSpeR P element vector to include a variety of unique restriction sites in the polylinker and also constructed two derivatives with the hsp70 promoter. As with the original pCaSpeR vector, these plasmids contain pUC sequences for propagation in bacteria and two P element ends flanking a white minigene and multiple cloning site. pCaSpeR 2 and pCaSpeR 3 contain a 39 bp oligonucleotide fragment in either of two orientations to introduce 5 new unique sites into the polylinker. pCaSpeR 4 contains the pW8 polylinker from the Gehring lab (Klemenz et al. (1987) Nuc. Acids Res. 15:3947-3959). pCaSpeR-hs is designed to allow heat-inducible expression of an inserted open reading frame. This P element contains an hsp70 promoter, multiple cloning site, and hsp70 3' trailer. The hsp70 3' trailer both reduces the accumulation of stable mRNA under non-heat shock conditions and allows the mRNA to induce to high levels following heat shock. pCaSpeR hs43 lacZ contains the lacZ reporter gene transcribed from a minimal promoter consisting of the hsp70 TATA box (deleted at -43) and leader sequences. It is derived from pCaSpeR 4 and is intended for testing enhancers and tissue- specific regulatory elements. The arrangement of restriction sites in these vectors is as follows, sites written in capital letters can be used for inserting DNA: pCaSpeR: HindIII/PSTI/SalI/XBAI/BAMHI/SmaI/SstI/ECORI pCaSpeR 2: HindIII/PSTI/SalI/XBAI/BAMHI/SmaI/SstI/STUI/XBAI/SSTII/NOTI/BG LII /HPAI/ECORI pCaSpeR 3: HindIII/PSTI/SalI/XBAI/BAMHI/SmaI/SstI/ECORI/HPAI/BGLII/NOTI/ SSTII/XBAI/STUI pCaSpeR 4: HindIII/PSTI/SalI/XHOI/STUI/XHOI/HPAI/SalI/XBAI/BAMHI/SPEI/SF II/ NOTI/SSTII/SmaI/KPNI/ECORI pCaSpeR-hs: hsp70 promoter/ECORI/HPAI/BGLII/NOTI/SSTII/XBAI/STUI/hsp70 polyA and 3' flanking pCaSpeR hs43 lacZ: ECORI/KpnI/SmaI/SSTII/NOTI/SFII/SPEI/BAMHI/XBAI/SalI/HPAI/XH OI/hs p43 promoter/PSTI/KpnI/SalI/SmaI/AUG lacZ *** CLONING OF LARGE SEGMENTS OF THE DROSOPHILA MELANOGASTER GENOME USING YEAST ARTIFICIAL CHROMOSOMES D. Filipp, G.L. Kogan, E.S. Belyaeva, B.A. Leibovitch, I.P. Arman, V.A. Gvozdev, Institute of Molecular Genetics, Acad. Sci. USSR, Moscow. A partial genomic library from the Batumi L stock of Drosophila melanogaster was constructed using yeast artificial chromosome vectors. The DNA was restricted with NotI and large fragments were inserted into the YAC5-vector (Burke, Carle, and Olson, 1987, Science 236: 806). The sizes of cloned DNA varied from 90 to 500 kb. 38 random clones were characterized by in situ hybridization to Batumi L salivary gland polytene chromosomes. Single euchromatic sites of hybridization were detected for 27 clones. The remaining 11 clones showed a main euchromatic site of hybridization and several additional sites scattered along the chromosomes (see list below). The clone DY 52 (500 kb) may have originated from the Y chromosome and DY 56 (90 kb) from the nucleolus. Any of the YAC clones described below are available to other investigators on request to Dr. G. L. Kogan. The inserts are, on average, 100 kb in length with a few that are 200 kb in length. Cytogenetic location of YAK hybridization: 1C+chromocenter, 5AB+chromocenter, 9C, 18DEF, 21CD, 28E, 34AB, 34C, 34D, 37AB, 44CD, 44D, 45E, 46AB+60BC, 46C, 46DEF+44D+98F+nucleolus, 47F, 48D, 52EF, 53A, 54A, 54CD, 54CD+nucleolus, 58EF, 59A+chromocenter, 60C, 61C+chromocenter+4th chromo, 61DE, 63E, 63E+1BC+62B, 65E+100A, 69F, 71C+chromocenter, 77E, 79E+76E+93A, 89A, 89B, 90DEF. *** EFFECT OF GROWTH MEDIUM ON ADH MAJOR/ADH MINOR ACTIVITY Leonard Borack[1], Richard Friedman[2], and William Sofer[2], [1]Dept. of Biol. Sci., Rutgers University, Newark, New Jersey 07102, USA. 201-648-5359. [2]Waksman Institute, Rutgers University, Piscataway, New Jersey 08854, USA. 201-932-3052, SOFER@MBCL.RUTGERS.EDU. Extracts of D. melanogaster third instar larvae of Canton S, WEP (Adh[F]), Adh[D], or Adh[fn6] somatic transformants, raised on Instant Drosophila Medium (Carolina Biological) show ADH[S], ADH[F] and ADH[D] minor form activity to be approximately 7% that of the corresponding ADH major form. Extracts of larvae raised on a Cornmeal-sucrose-molasses medium show ADH[S], ADH[F] and ADH[D] minor form activity at approximately 20% that of the corresponding ADH major form. After electrophoresis, ADH[F] major and ADH[S] minor are seen to co-migrate, as do ADH[D] major and ADH[F] minor. These differences in the percentage of minor band activity can affect calculations of ADH major form activity when two or more alleles are assayed after electrophoresis in the same lane. ***