Method for the controlled synthesis of polynucleotide...

C - Chemistry – Metallurgy – 12 – N

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Details

C12N 15/10 (2006.01) C07H 21/00 (2006.01) C12N 15/11 (2006.01) C12Q 1/68 (2006.01) G01N 33/554 (2006.01)

Patent

CA 2208693

A method to obtain selected individual polynucleotides or mixtures thereof each of which encodes a peptide and at least one polynucleotide of the mixture encodes a peptide having a target property. The polynucleotides of the invention present in the mixture in detectable, retrievable, and clonable amounts are expressed in a host organism for screening for the target activity. The invention features the ability to synthesize controlled random polynucleotides to produce a predetermined mixture of polynucleotides and to avoid synthesis of a stop codon by adjusting the proportions into which the synthesis pool is subdivided and by adjusting the proportions of activated nucleotides added at each coupling step. A polynucleotide encoding a peptide having a target property can be selected and sequenced to deduce the amino acid sequence of the peptide.

Méthode pour obtenir des polynucléotides individuels sélectionnés ou leurs mélanges; chacun d'entre eux code un peptide et au moins un polynucléotide du mélange code un peptide possédant une propriété cible. Les polynucléotides de l'invention, présents dans le mélange en quantités décelables, récupérables et clonables, sont exprimés chez un organisme hôte pour le dépistage de l'activité cible. L'invention permet de synthétiser des polynucléotides statistiques contrôlés et d'obtenir ainsi un mélange de polynucléotides déterminé préalablement, en évitant la synthèse d'un codon d'arrêt par ajustement des proportions de la subdivision du bassin de synthèse ainsi que des proportions de nucléotides activés ajoutés à chaque étape de couplage. Un polypeptide codant un peptide avec propriété cible peut être choisi et séquencé pour déduire la séquence d'amino-acides du peptide.

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