Construction of normalized cdna libraries from eucaryotic cells

Details Construction of normalized cdna libraries from eucaryotic cells Construction of normalized cdna libraries from eucaryotic cells

C - Chemistry, Metallurgy

12

N

C12N 15/10 (2006.01) C12N 15/62 (2006.01)

Patent

CA 2339041

A new technology is described that allows for the rapid and efficient construction of complex cDNA libraries from cultured eukaryotic cells. The technology exploits eukaryotic biology by using transgenic constructs that have been nonspecifically inserted into the genome to facilitate the expression of nuclear genes as fusion transcripts. The invention further allows one to specifically subclone the corresponding fusion transcripts into highly complex cDNA libraries. The libraries are easily characterized by molecular analysis techniques such as hybridization, and individual clones can be directly sequenced to generate a sequence database of the cellular portion of the fusion transcripts.

L'invention concerne une nouvelle technologie permettant la construction rapide et efficace de bibliothèques d'ADNc complexes, à partir de cellules eucaryotes cultivées. Cette technologie exploite la biologie euraryote au moyen de constructions transgéniques qui n'ont pas été spécifiquement insérées dans le génome, afin de faciliter l'expression des gènes nucléaires comme transcrits de fusion. L'invention permet également de sous-cloner spécifiquement les transcrits de fusion correspondants en bibliothèques d'ADNc très complexes. Ces bibliothèques sont facilement caractérisées par des techniques d'analyse moléculaire telles que l'hybridation, et des clones individuels peuvent être directement séquencés, afin de produire une base de données de séquences de la partie cellulaire des transcrits de fusion.

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