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1. WO2000017342 - INTRON-MEDIATED RECOMBINANT TECHNIQUES AND REAGENTS

Publication Number WO/2000/017342
Publication Date 30.03.2000
International Application No. PCT/US1999/021929
International Filing Date 21.09.1999
Chapter 2 Demand Filed 29.03.2000
IPC
C12N 15/10 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
CPC
C12N 15/10
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
C12N 15/63
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
Applicants
  • TRUSTEES OF BOSTON UNIVERSITY [US]/[US]
Inventors
  • JARRELL, Kevin, A.
  • MULCHEEVA, Svetlana
  • DONAHUE, William
Agents
  • NUGENT, Elizabeth, E.
  • JARRELL, Brenda, Herschbach
Priority Data
60/101,32821.09.1998US
null20.09.1999US
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) INTRON-MEDIATED RECOMBINANT TECHNIQUES AND REAGENTS
(FR) PROCEDES ET REACTIFS DE RECOMBINAISON A MEDIATION INTRONIQUE
Abstract
(EN)
The present invention makes available methods and reagents for novel manipulation of nucleic acids. As described herein, the present invention makes use of the ability of intronic sequences, such as derived from group I, group II, or nuclear pre-mRNA introns, to mediate specific cleavage and ligation of discontinuous nucleic acid molecules. For example, novel genes and gene products can be generated by admixing nucleic acid constructs which comprise exon nucleic acid sequences flanked by intron sequences that can direct trans-splicing of the exon sequences to each other. The flanking intronic sequences can, by intermolecular complementation, form a reactive complex which promotes the transesterification reactions necessary to cause the ligation of discontinuous nucleic acid sequences to one another, and thereby generate a recombinant gene comprising the ligated exons.
(FR)
L'invention concerne des procédés et des réactifs permettant une manipulation nouvelle des acides nucléiques. On utilise l'aptitude des séquences introniques (par exemple, dérivées d'introns des groupes I, II ou de type nucléaire pré-ARNm) à assurer la médiation d'opérations spécifiques de clivage et de ligature de molécules discontinues d'acides nucléiques. Ainsi, il est possible d'élaborer de nouveaux gènes et produits géniques par admixtion de constructions d'acides nucléiques comprenant des séquences d'acides nucléiques exoniques flanquées de séquences introniques capables d'orienter mutuellement l'une vers l'autre les séquences exoniques en trans-épissage. Par complémentation intermoléculaire, les séquences introniques flanquantes peuvent former un complexe réactif qui favorise les réactions de transestérification requises pour la ligature de séquences nucléotidiques discontinues, moyennant quoi il est possible d'élaborer un gène de recombinaison renfermant les exons ligaturés.
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