In vitro transposition of artificial transposons

Details In vitro transposition of artificial transposons In vitro transposition of artificial transposons

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C12N 15/10 (2006.01) C12N 15/11 (2006.01) C12N 15/90 (2006.01)

Patent

CA 2161767

We have developed efficient methods of creating artificial transposons and inserting these transposons into DNA targets in vitro, primarily for the purpose of mapping and sequencing DNA. A target DNA has been engineered to convert virtually any DNA sequence, or combination of sequences, into an artificial transposon; hence, custom transposons containing any desired feature can be easily designed and constructed. Such transposons are then efficiently inserted into DNA targets, in vitro, using the integrase activity present in yeast Ty1 virus-like particles. Primers complementary to the transposon termini can be used to sequence DNA flanking any transposon insertion.

On a développé des procédés efficaces de création de transposons artificiels et d'insertion de ces transposons dans des cibles d'ADN in vitro, principalement afin de cartographier et de séquencer l'ADN. On a mis au point génétiquement un ADN cible afin de convertir virtuellement n'importe quelle séquence d'ADN, ou combinaison de séquences, dans un transposon artificiel, à partir de quoi on peut concevoir et construire facilement des transposons adaptés en vue d'une utilisation particulière, contenant n'importe quelle caractéristique voulue. On insère ensuite efficacement lesdits transposons dans des cibles d'ADN, in vitro, à l'aide de l'activité intégrase présente dans des particules de levure Ty1 de type viral. On peut utiliser des amorces complémentaires aux terminaisons des transposons pour séquencer l'ADN bordant n'importe quelle insertion de transposon.

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