"doping" in walk-through mutagenesis

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/68 (2006.01) C12N 15/10 (2006.01) C12N 15/63 (2006.01) C12P 21/00 (2006.01)

Patent

CA 2482769

A method of walk-through mutagenesis of a nucleic acid encoding a prototype polypeptide of interest, is described, the method comprising selecting a predetermined amino acid and one or more target regions of the polypeptide, and synthesizing a mixture of oligonucleotides containing at each sequence position in the target region, either a prototype nucleotide that is required for synthesis of the prototype amino acid of the polypeptide, or a predetermined nucleotide that is required for synthesis of the predetermined amino acid, in which during the synthesis, the ratio of available prototype nucleotides, to available predetermined nucleotides, is greater than 1:1.

L'invention concerne un procédé de mutagenèse traversante d'un acide nucléique codant un polypeptide prototype d'intérêt. Ledit procédé comprend le fait de sélectionner un aminoacide prédéterminé et au moins une région cible du polypeptide et de synthétiser un mélange d'oligonucléotides contenant à chaque position de séquence dans la zone cible, soit un nucléotide prototype requis pour la synthèse de l'aminoacide prototype du polypeptide, soit un nucléotide prédéterminé requis pour la synthèse de l'aminoacide prédéterminé, dans lequel pendant la synthèse, le rapport des nucléotides prototypes disponibles aux nucléotides prédéterminés disponibles est supérieur à 1 :1.

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