METHOD FOR OBTAINING IN VITRO RECOMBINED POLYNUCLEOTIDE SEQUENCES, SEQUENCE BANKS AND RESULTING SEQUENCES

• Main Authors: DUPRET, DANIEL, MASSON, JEAN-MICHEL, LEFEVRE, FABRICE
• Format: Patent
• Language: English; French
• Published: 24.02.2000
• Edition: 7
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TESTING; VINEGAR; WINE

La présente invention a pour objet un procédé de production in vitro de séquences polynucléotidiques recombinées à partir d'une banque de séquences polynucléotidiques, caractérisé en ce qu'il comprend les étapes suivantes: ( a) la fragmentation d'une banque initiale de séquences polynucléotidiques doubl e- brins; (b) la dénaturation des fragments issus de l'étape (a) éventuellement en présence d'une ou plusieurs matrice(s) d'assemblage; (c) l'hybridation desdits fragments avec une ou plusieurs matrice(s) d'assemblage si celle(s)- ci n'est (ne sont) pas présente(s) à l'étape (b); (d) la ligation desdits fragments; (e) éventuellement le clonage des séquences polynucléotidiques recombinées, et la sélection des séquences polynucléotidiques recombinées, e t la sélection des séquences polynucléotidiques présentant des propriétés avantageuses par rapport à une ou plusieurs séquences de référence. A method (I) for producing recombined polynucleotide sequences from a library of double-stranded polynucleotide sequences is new and comprises fragmenting and denaturing library sequences, hybridizing them with assembly matrices and selecting sequences with requires properties.. Method (I) for producing recombined polynucleotide sequences from a library of double-stranded polynucleotide sequences is new and comprises: (1) fragmenting the library sequences; (2) denaturing the fragments, optionally in the presence of one or more assembly matrices; (3) hybridizing the fragments with one or more assembly matrices, if these were not present in step (b); (4) ligating the fragments to obtain recombined sequences; and (5) selecting recombined sequences having advantageous properties compared with one or more reference sequences. Independent claims are also included for the following: (1) a recombined polynucleotide sequence obtained and selected by method (I); (2) a vector containing the sequence of (1); (3) a host cell transformed with the sequence of (1) or the vector of (2); (4) a protein encoded by the sequence of (1); (5) a library of polynucleotide sequences as in (1), vectors as in (2), cells as in (3) or proteins as in (4).