Selection subtraction approach to gene identification

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 5/00 (2006.01) C12N 15/10 (2006.01) C12N 15/13 (2006.01) C12N 15/867 (2006.01) C12Q 1/68 (2006.01) G01N 33/53 (2006.01)

Patent

CA 2323633

The present invention provides a method of creating expression libraries by marking desired clones with a unique tag. An expression library including individual clones marked with a unique tag sequence is also provided. Also provided is a method of isolating genetic elements associated with complete or partial cell growth suppression or cell death by isolating clones from an expression library which are lost in the cell population. A method of selection subtraction is provided by tagging a clone in an expression library, cloning the tagged clone into a vector, delivering the tagged clone to a target cell, and comparing tags before and after selection whereby toxic genes and the attached tags disappear.

La présente invention porte sur un procédé de création de banques d'expression, procédé consistant à marquer des clones désirés avec une étiquette unique. L'invention porte également sur une banque d'expression comprenant des clones individuels marqués d'une séquence d'étiquette unique; sur un procédé visant à isoler des éléments génétiques associés à la suppression de la prolifération cellulaire ou mort cellulaire totale ou partielle, des clones, perdus dans une population cellulaire, étant ainsi isolés d'une banque d'expression. L'invention porte, d'autre part, sur un procédé de sélection-soustraction qui consiste à étiqueter un clone dans une banque d'expression, cloner le clone étiqueté dans un vecteur, acheminer le clone étiqueté vers une cellule cible, et comparer les étiquettes avant et après sélection pour que disparaissent les gènes toxiques et leurs étiquettes.

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