New method for the selection of clones of an expression...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12N 15/10 (2006.01) C12N 15/64 (2006.01)

Patent

CA 2329986

The present invention relates to a novel method for the selection of clones of an expression library. The method is characterized by the step of analyzing for the expression of at least one (poly)peptide, such as a tag expressed in frame with a recombinant insert of said expression library, wherein said clones of said expression library are arranged in arrayed form. Screening for the expression of said (poly)peptide allows for the rearraying of clones that express the recombinant insert. Upon rearraying said clones, the library will now comprise clones having inserts in the correct reading frame to a very high degree. Optionally, the library is additionally screened for desired inserts either by contacting the expressed library with a ligand specific for the expression product or by hybridizing or fingerprinting the inserts with a nucleic acid probe.

L'invention concerne un nouveau procédé de sélection de clones dans une banque d'expression. Ce procédé est caractérisé par l'identification de l'expression d'au moins un (poly)peptide tel qu'un marqueur, exprimé de pair avec un insert recombinant provenant de cette banque d'expression, les clones composant la banque d'expression étant disposés sous forme d'une matrice. La recherche de l'expression de ce (poly)peptide permet le réarrangement des clones exprimant l'insert recombinant. Après ce réarrangement, le banque comprend des clones comprenant en très grande partie des inserts dans la phase de lecture correcte. On peut en outre rechercher des inserts désirés dans la banque soit en mettant en contact la banque d'expression avec un ligand spécifique du produit d'expression, soit en utilisant une sonde d'acide nucléique pour hybrider les inserts ou établir une carte peptidique de ces derniers.

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