Title:
METHODS AND MATERIALS FOR THE RAPID AND HIGH VOLUME PRODUCTION OF A GENE KNOCK-OUT LIBRARY IN AN ORGANISM
Document Type and Number:
WIPO Patent Application WO2000055346
Kind Code:
A3
Abstract:
The present invention relates to a method for facilitating site directed homologous recombination in an organism to produce mutants comprising: 1) constructing a large insert vector library comprising one or more large insert vectors; 2) constructing a second vector comprising a transposable element; 3) incubating the product of step 1) with the product of step 2) in the presence of a transposase specific for the inverted repeat sequences on the plasmid vector; 4) amplifying the disrupted large insert vector resulting from step 3); 5) introducing the disrupted large insert vector produced by step 4) into a target host cell; and 6) selecting for successful homologous recombination produced by step 5) with the second selectable marker.
Inventors:
HAMER JOHN E (US)
HAMER LISBETH (US)
HAMER LISBETH (US)
Application Number:
PCT/US2000/007317
Publication Date:
February 15, 2001
Filing Date:
March 16, 2000
Export Citation:
Assignee:
PARADIGM GENETICS INC (US)
HAMER JOHN E (US)
HAMER LISBETH (US)
HAMER JOHN E (US)
HAMER LISBETH (US)
International Classes:
C12N15/09; C12N9/22; C12N15/10; C12N15/90; (IPC1-7): C12N15/90; C12N9/22; C12N15/10
Domestic Patent References:
| WO1998056903A1 | 1998-12-17 | |||
| WO1998010077A1 | 1998-03-12 | |||
| WO1997029202A1 | 1997-08-14 | |||
| WO1999009817A1 | 1999-03-04 | |||
| WO1999007871A1 | 1999-02-18 |
Foreign References:
| US5102797A | 1992-04-07 |
Other References:
WESTPHAL CHRISTOPH HEINER ET AL: "Transposon-generated 'knock-out' and 'knock-in' gene-targeting constructs for use in mice.", CURRENT BIOLOGY, vol. 7, no. 7, 1997, pages 530 - 533, XP000938425, ISSN: 0960-9822
LAMPE DAVID J ET AL: "A purified mariner transposase is sufficient to mediate transposition in vitro.", EMBO (EUROPEAN MOLECULAR BIOLOGY ORGANIZATION) JOURNAL, vol. 15, no. 19, 1996, pages 5470 - 5479, XP002145182, ISSN: 0261-4189
LAMPE DAVID J ET AL: "Factors affecting transposition of the Himar1 mariner transposon in vitro.", GENETICS, vol. 149, no. 1, May 1998 (1998-05-01), pages 179 - 187, XP002145183, ISSN: 0016-6731
NISHIMURA MARIE ET AL: "Construction of a BAC library of the rice blast fungus Magnaporthe grisea and finding specific genome regions in which its transposons tend to cluster.", BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, vol. 62, no. 8, August 1998 (1998-08-01), pages 1515 - 1521, XP002145184, ISSN: 0916-8451
ZHANG L ET AL: "THE HIMAR1 MARINER TRANSPOSASE CLONED IN A RECOMBINANT ADENOVIRUS VECTOR IS FUNCTIONAL IN MAMMALIAN CELLS", NUCLEIC ACIDS RESEARCH,GB,OXFORD UNIVERSITY PRESS, SURREY, vol. 26, no. 16, August 1998 (1998-08-01), pages 3687 - 3693, XP000700201, ISSN: 0305-1048
RUBIN ERIC J ET AL: "In vivo transposition of mariner-based elements in enteric bacteria and mycobacteria.", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES, vol. 96, no. 4, 16 February 1999 (1999-02-16), Feb. 16, 1999, pages 1645 - 1650, XP002145185, ISSN: 0027-8424
ADACHI KIICHI AND J.E. HAMER: "Divergent cAMP signaling pathways regulate growth and pathogenesis in the rice blast fungus Magnaporthe grisea.", PLANT CELL, vol. 10, no. 8, August 1998 (1998-08-01), pages 1361 - 1373, XP002145186, ISSN: 1040-4651
S.E. DEVINE ET AL.: "A transposon-based strategy for sequencing repetitive DNA in eucaryotic genomes", GENOME RESEARCH, vol. 7, 1997, CSH LABORATORY PRESS, pages 551 - 563, XP002145187
LAMPE DAVID J ET AL: "A purified mariner transposase is sufficient to mediate transposition in vitro.", EMBO (EUROPEAN MOLECULAR BIOLOGY ORGANIZATION) JOURNAL, vol. 15, no. 19, 1996, pages 5470 - 5479, XP002145182, ISSN: 0261-4189
LAMPE DAVID J ET AL: "Factors affecting transposition of the Himar1 mariner transposon in vitro.", GENETICS, vol. 149, no. 1, May 1998 (1998-05-01), pages 179 - 187, XP002145183, ISSN: 0016-6731
NISHIMURA MARIE ET AL: "Construction of a BAC library of the rice blast fungus Magnaporthe grisea and finding specific genome regions in which its transposons tend to cluster.", BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, vol. 62, no. 8, August 1998 (1998-08-01), pages 1515 - 1521, XP002145184, ISSN: 0916-8451
ZHANG L ET AL: "THE HIMAR1 MARINER TRANSPOSASE CLONED IN A RECOMBINANT ADENOVIRUS VECTOR IS FUNCTIONAL IN MAMMALIAN CELLS", NUCLEIC ACIDS RESEARCH,GB,OXFORD UNIVERSITY PRESS, SURREY, vol. 26, no. 16, August 1998 (1998-08-01), pages 3687 - 3693, XP000700201, ISSN: 0305-1048
RUBIN ERIC J ET AL: "In vivo transposition of mariner-based elements in enteric bacteria and mycobacteria.", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES, vol. 96, no. 4, 16 February 1999 (1999-02-16), Feb. 16, 1999, pages 1645 - 1650, XP002145185, ISSN: 0027-8424
ADACHI KIICHI AND J.E. HAMER: "Divergent cAMP signaling pathways regulate growth and pathogenesis in the rice blast fungus Magnaporthe grisea.", PLANT CELL, vol. 10, no. 8, August 1998 (1998-08-01), pages 1361 - 1373, XP002145186, ISSN: 1040-4651
S.E. DEVINE ET AL.: "A transposon-based strategy for sequencing repetitive DNA in eucaryotic genomes", GENOME RESEARCH, vol. 7, 1997, CSH LABORATORY PRESS, pages 551 - 563, XP002145187
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