In vitro recombination of polynucleotide fragments to obtain sequences with improved properties involves ligation on an assembly matrix

• Main Authors: DUPRET DANIEL, LEFEVRE FABRICE, MASSON JEAN MICHEL
• Format: Patent
• Language: English; French
• Published: 18.02.2000
• 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

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). La présente invention se rapporte à une méthode de production in vitro de séquences polynucléotidiques recombinées comprenant les étapes suivantes : (a) la fragmentation d'une banque initiale de séquences polynucléotidiques double-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 en présence d'une ou plusieurs matrice (s) d'assemblage si celle (s) -ci n'est (ne sont) pas présente (nt) à l'étape (b), (d) la ligation desdits fragments, (e) le clonage des séquences polynucléotidiques recombinées.