COLD SPRING HARBOR KEY LAB MANUALS from LABORATORY PRESS MOLECULAR CLONING A LABORATORY MANUAL, SECOND EDITION J. Sambrook, University of Texas Southwestern Medical Center; E.F. Fritsch, Genetics Institute; T. Maniatis, Harvard University "The [second edition] of Molecular Cloning is indeed an impressive achievement. Its growth in size alone bears witness to the explosion in molecular techniques .... This ex- pansion reflects more comprehensive coverage of topics previously included in the first edition as well as the addition of new chapters, such as those on oligonucleotide probes and mutagenesis, in vitro amplification by the polymerase chain reaction, expression of cloned genes in Escherichia coil and mammalian cells, and analysis of proteins ex- pressed from cloned genes. Particularly noteworthy are the sections dealing with con- struction of complementary DNA and genomic libraries. Current tendencies to 'clone by kit' and 'clone by phone' aside, these sections provide thoughtful accounts of different approaches with a balanced view of their inherent strengths and weaknesses. As with the previous edition, the protocols are presented in a pleasing, easy-to-read format .... Molecular Cloning is of truly exceptional value. By virtue of its heritage and modest cost, it is likely to be more widely used by students at all stages of development."_Natur e 1989, 1659 pp., illus., appendices, bibliography, index ISBN 0-87969-309-6 Plastic comb binding, 3-book set $125 A SHORT COURSE IN BACTERIAL GENETICS A Laboratory Manual and Handbook for Escherichia coil and Related Bacteria Jeffrey H. Miller, University of California, Los Angeles A long-awaited sequel to Experiments in Molecular Genetics, the two-part volume A Short Course in Bacteria/Genetics: A Laboratory Manual and Handbook for Escherichia coil and Related Bacteria is essential for all those doing genetic or recombinant DNA work with E. coil or similar organisms. The Manual includes 34 detailed experiments with step-by-step protocols and easy-to-follow diagrams that demonstrate major concepts in experimental bacterial genetics. The experiments cover the essential points of mutagenesis, gene trans- fer, transposable elements, and gene fusions and are accomplished with a set of 44 bac- terial strains. The strains are prepared in the author's laboratory and sold by Cold Spring Harbor Laboratory Press as a kit. The Handbook summarizes important information about E. co/i, its genes and proteins, phage and plasmid vectors, transposable elements (including relevant sequence informa- tion), the current detailed genetic map, and the complete restriction maps of several cosmid and phage libraries of the entire genome. 1992, 876 pp., illus., color plates, bibliography, index ISBN 0-87969-349-5 Plastic comb bound Manual and Paperback Handbook $110 A Short Course in Bacterial Genetics Strain Kit. A kit of 44 bacterial strains and phage lysates is available through Cold Spring Harbor Laboratory Press. Prepared and tested in the author's laboratory at UCLA, the strains are used in the set of experiments described in A Short Course in Bacteria/Genetics and are valuable for many additional experiments. (Please give your phone number when ordering and allow 6 weeks for delivery). $350 ISBN 0-87969..400-9 ANTIBODIES A LABORATORY MANUAL Ed Harlow, Cold Spring Harbor Laboratory; David Lane,/mperial Cancer Research Fund Laboratories Antibodies aims "to put current immunological methodology into the hands of non- specialists. Happily, Harlow and Lane have succeeded admirably and this manual seems certain also to gain a place on the laboratory bench (too useful to remain on the bookshelf) of a wide variety of biologists-even immunologists. The volume opens with four introductory chapters summarizing key features of the immune response; these provide a succinct overview of the field and are very well written. The scope of the rest of the book includes techniques of immunization and bleeding, monoclonal antibodies and hybridomas, protein labelling, immunoassays (mostly those using solid supports), immunoprecipitation of cellular antigens, immunoblotting and immunoaffinity purifica- tion .... The methods are presented as easy-to-follow '1-2-3 protocols,' and assume rela- tively little experience. Rationales for procedures are often presented, which helps to remove some of the mystique that has been associated with immunological methodol- ogy." -Nature 1988, 726 pp., illus., appendices, bibliography, index ISBN 0-87969-314-2 Plastic comb binding $70 R~=~dr=r R~rvir.r= Nn. 195
Transcript
COLD SPRING HARBOR K E Y LAB M A N U A L S from LABORATORY PRESS
MOLECULAR CLONING A LABORATORY MANUAL, SECOND EDITION J. Sambrook, University of Texas Southwestern Medical Center; E.F. Fritsch, Genetics Institute; T. Maniatis, Harvard University
"The [second edition] of Molecular Cloning is indeed an impressive achievement. Its growth in size alone bears witness to the explosion in molecular techniques .... This ex- pansion reflects more comprehensive coverage of topics previously included in the first edition as well as the addition of new chapters, such as those on oligonucleotide probes and mutagenesis, in vitro amplification by the polymerase chain reaction, expression of cloned genes in Escherichia coil and mammalian cells, and analysis of proteins ex- pressed from cloned genes. Particularly noteworthy are the sections dealing with con- struction of complementary DNA and genomic libraries. Current tendencies to 'clone by kit' and 'clone by phone' aside, these sections provide thoughtful accounts of different approaches with a balanced view of their inherent strengths and weaknesses. As with the previous edition, the protocols are presented in a pleasing, easy-to-read format .... Molecular Cloning is of truly exceptional value. By virtue of its heritage and modest cost, it is likely to be more widely used by students at all stages of development."_Natur e
1989, 1659 pp., illus., appendices, bibliography, index ISBN 0-87969-309-6 Plastic comb binding, 3-book set $125
A SHORT COURSE IN BACTERIAL GENETICS A Laboratory Manual and Handbook for Escherichia coil and Related Bacteria Jeffrey H. Miller, University of California, Los Angeles
A long-awaited sequel to Experiments in Molecular Genetics, the two-part volume A Short Course in Bacteria/Genetics: A Laboratory Manual and Handbook for Escherichia coil and Related Bacteria is essential for all those doing genetic or recombinant DNA work with E. coil or similar organisms. The Manual includes 34 detailed experiments with step-by-step protocols and easy-to-follow diagrams that demonstrate major concepts in experimental bacterial genetics. The experiments cover the essential points of mutagenesis, gene trans- fer, transposable elements, and gene fusions and are accomplished with a set of 44 bac- terial strains. The strains are prepared in the author's laboratory and sold by Cold Spring Harbor Laboratory Press as a kit.
The Handbook summarizes important information about E. co/i, its genes and proteins, phage and plasmid vectors, transposable elements (including relevant sequence informa- tion), the current detailed genetic map, and the complete restriction maps of several cosmid and phage libraries of the entire genome. 1992, 876 pp., illus., color plates, bibliography, index ISBN 0-87969-349-5 Plastic comb bound Manual and Paperback Handbook $110
A Short Course in Bacterial Genetics Strain Kit. A kit of 44 bacterial strains and phage lysates is available through Cold Spring Harbor Laboratory Press. Prepared and tested in the author's laboratory at UCLA, the strains are used in the set of experiments described in A Short Course in Bacteria/Genetics and are valuable for many additional experiments. (Please give your phone number when ordering and allow 6 weeks for delivery). $350 ISBN 0-87969..400-9
ANTIBODIES A LABORATORY MANUAL Ed Harlow, Cold Spring Harbor Laboratory; David Lane,/mperial Cancer Research Fund Laboratories
Antibodies aims "to put current immunological methodology into the hands of non- specialists. Happily, Harlow and Lane have succeeded admirably and this manual seems certain also to gain a place on the laboratory bench (too useful to remain on the bookshelf) of a wide variety of biologists-even immunologists. The volume opens with four introductory chapters summarizing key features of the immune response; these provide a succinct overview of the field and are very well written. The scope of the rest of the book includes techniques of immunization and bleeding, monoclonal antibodies and hybridomas, protein labelling, immunoassays (mostly those using solid supports), immunoprecipitation of cellular antigens, immunoblotting and immunoaffinity purifica- tion .... The methods are presented as easy-to-follow '1-2-3 protocols,' and assume rela- tively little experience. Rationales for procedures are often presented, which helps to remove some of the mystique that has been associated with immunological methodol- ogy." -Nature 1988, 726 pp., illus., appendices, bibliography, index ISBN 0-87969-314-2 Plastic comb binding $70
R~=~dr=r R~rvir.r= Nn. 195
GENES
DEVELOPMENT
V O L U M E 8 N U M B E R 15 PAGES 1739--1866 A U G U S T I, 1994
EDITORIAL BOARD
J. Adams (Melboume, Australia) J. Beckwith (Boston, USA) T. Cech (Boulder, USA) P. Chambon (Strasbourg, France) N.-H. Chua (New York, USA) E. Coen {Norwich, UK) J. Coffin (Boston, USA) S. Courtneidge (Heidelberg, FRG) E. De Robertis (Los Angeles, USA) R. Evans (La Jolla, USA) G. Fink (Cambridge, USA) P. Goodfellow (Cambridge, UK) S. Gottesman (Bethesda, USA} T. Graf (Heidelberg, FRG) C. Gross (San Francisco, USA) R. Grosschedl (San Francisco, USA) F. Grosveld (Rotterdam, The Netherlands) M. Groudine (Seattle, USA) L. Guarente (Cambridge, USA) R. Harland (Berkeley, USA) E. Harlow (Charlestown, USA) W. Herr (Cold Spring Harbor, USA) J. Hodgkin (Cambridge, UK) R. Horvitz (Cambridge, USA) P. Ingham (London, UK) T. lessell (New York, USA)
N. Jones (London, UK) J. Kadonaga (La Jolla, USA) R. Lehmann (Cambridge, USA) M. Levine (San Diego, USA) D. Livingston (Boston, USA) R. Losick (Cambridge, USA) J. Manley {New York, USA) W. McGinnis {New Haven, USA) S. McKnight (South San Francisco, USA) A. McMahon (Cambridge, USA) P. Nurse (London, UK) C. Nfisslein-Volhard (Tubingen, FRG) R. Palmiter (Seattle, USA) L. Parada (Dallas, USA) C. Prives (New York, USA) G. Rubin (Berkeley, USA) U. Schibler (Geneva, Switzerland) D. Solter {Freiburg, FRG) P. Soriano (Seattle, USA) J. Steitz (New Haven, USA) T. Taniguchi (Osaka, Japan) S. Tilghman (Princeton, USA) R. Tjian (Berkeley, USA) E. Wagner (Vienna, Austria) V. Walbot (Stanford, USA) M. Wiglet (Cold Spring Harbor, USA)
Editors T. Grodzicker (Cold Spring Harbor) N. Hastie (Edinburgh)
Managing Editor J. Cuddihy (Cold Spring Harbor)
Editorial~Production N. Dumser, Technical Editor V. Nicolette, Production Editor L. Olsewski, Editorial Secretary
EDITORIAL OFFICES
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GENES & DEVELOPMENT {ISSN 0890-9369 is published semimonthly for $450 (institutional), $115 (individual making personal payment), $159 (Genet- ical Society of Great Britain members) by Cold Spring Harbor Laboratory Press, Bungtown Road, Cold Spring Harbor, New York 11724, in associ- ation with the Genetical Society of Great Britain. Second-class postage is paid at Cold Spring Harbor and additional mailing offices. POSTMAS- TER: Send address changes to Cold Spring Harbor Laboratory Press, POB 100, 1 Bungtown Rd., Cold Spring Harbor, New York 11724-2203. Subscription Price Orders may be sent to Cold Spring Harbor Labora- tory Press, Fulfillment Department, 10 Skyline Drive, Plainview, NY 11803-9729. Telephone: Continental US except NY State, 1-800-843- 4388; all other locations, 516-349-1930/1931/1932. FAX 516-349-1946. Volume 8, 1994, $450, U.S. institutional; $540, R.O.W. institutional. Personal subscription rate: $115, U.S.; $205, R.O.W. Price includes sur- face postage for U.S. and airlift for R.O.W. Genetical Society members, $159.00. All subscriptions are entered for the calendar year and must be prepaid. Personal subscriptions must be prepaid by personal check, credit card, or money order. All checks must be for US dollars and drawn on a US bank. Genetical Society members may also subscribe by check, payable to the Genetical Society, for s (includes airlift). Send to: Dr.
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Claims for missing issues must be received within 4 months of issue date. Change of address Please enclose recent mailing label with address change; allow 4 weeks. Advertising To advertise in Genes & Development, contact Nancy Kuhle, Advertising Manager, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York 11724-2203i telephone 516-367-8351. Photo Copy Authorization to photocopy items for internal or personal use, or the internal or personal use of specific clients, is granted by Cold Spring Harbor Laboratory Press for libraries and other users registered with the Copyright Clearance Center {CCC) Transactional Reporting Service, provided that the base fee of $5.00 per copy is paid directly to CCC, 21 Congress St., Salem, MA 01970 (0890-9369/94 $5.00 + 0.). This consent does not extend to other kinds of copying, such as copying for general distribution for advertising or promotional purposes, for creating new collective works, or for resale. Copyright �9 1994 by Cold Spring Harbor Laboratory Press
Contents GENES & DEVELOPMENT August 1, 1994
Research papers
Regulation of mdm2 expression by p53: alternative promoters produce transcripts with nonidentical translation potential Yaacov Barak, Eyal Gottlieb, Tamar Juven-Gershon, and Moshe Oren
1739
p21-containing cyclin kinases exist in both active and inactive states Hui Zhang, Gregory J. Hannon, and David Beach
1750
Collaboration of G1 cyclins in the functional inactivation of the retinoblastoma protein Masanori Hatakeyama, Julie A. Brill, Gerald R. Fink, and Robert A. Weinberg
1759
Differential regulation of E2F trans-activation by cyclin/cdk2 complexes Brian David Dynlacht, Osvaldo Flores, Jacqueline A. Lees, and Ed Harlow
1772
Distinct morphogenetic functions of similar small GTPases: Drosophila Dracl is involved in axonal outgrowth and myoblast fusion Liqun Luo, Y. Joyce Liao, Lily Yeh Jan, and Yuh Nung Jan
1787
Ras/MAP kinase-dependent and -independent signaling pathways target distinct ternary complex factors Robert A. Hipskind, Dirk B6scher, Alfred Nordheim, and Manuela Baccarini
1803
The influenza virus NS1 protein: a novel inhibitor of pre-mRNA splicing Yuan Lu, Xiao-Yan Qian, and Robert M. Krug
1817
Short-range repression permits multiple enhancers to function autonomously within a complex promoter Susan Gray, Paul Szymanski, and Michael Levine
1829
A ~r s4 transcriptional activator also functions as a pole-specific repressor in Caulobacter James A. Wingrove and James Gober
1839
Molecular and genetic characterization of GABP~ Fabienne Charles de la Brousse, Edward H. Birkenmeier, David S. King, Lucy B. Rowe, and Stephen L. McKnight
1853
Cover Lateral (top} and ventral {bottom) views of transgenic Drosophila embryos carrying a synthetic promoter element (diagram) containing bicoid, dorsal, and snail binding sites. The resulting expression pattern represents the combined action of these three regulatory proteins based on their known spatial distributions. (For details, see Gray et al., p. 1829.)
