Reassortment by fragment ligation

Details Reassortment by fragment ligation Reassortment by fragment ligation

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2660705

The present invention provides methods for preparing polynucleotide variants. In one aspect of the invention, the method comprises: a) exposing a pool of two or more related polynucleotides to at least one nicking enzyme, where at least some of the polynucleotides are partially and/or fully double stranded, b) removing, and/or inhibiting the activity of, the at least one nicking enzyme, c) denaturing the polynucleotides, d) allowing the denatured polynucleotides to form at least partially double stranded polynucleotides, e) exposing the double stranded polynucleotides formed in step d) to a ligase. Also provided are methods of making a polynucleotide and/or polypeptide having a desired property.

La présente invention concerne des procédés destinés à obtenir des variants polynucléotidiques. Dans un aspect de l'invention, le procédé comprend les étapes consistant : a) à exposer un ensemble de deux polynucléotides apparentés ou plus à au moins une enzyme de coupure, au moins certains des polynucléotides étant partiellement et/ou entièrement à double brin, b) à supprimer et/ou à inhiber l'activité de ladite ou desdites enzymes de coupure, c) à dénaturer les polynucléotides, d) à permettre aux polynucléotides dénaturés de constituer des polynucléotides au moins partiellement à double brin, et enfin, e) à exposer les polynucléotides à double brin constitués lors de l'étape d) à une ligase. La présente invention concerne également des procédés destinés à fabriquer un polynucléotide et/ou un polypeptide ayant une propriété désirée.

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