| 主权项 |
1. A method of producing an organism with a new or altered phenotype, comprising:
(a) introducing a composition comprising a plurality of random in-frame fusion polynucleotides having sequences different from each other into an organism to produce one or more transformed organisms, each random in-frame fusion polynucleotide comprising at least two full-length open reading frames, one designated as a 5′full-length open reading frame and one designated as a 3′ full-length open reading frame, the 5′ full-length open reading frame being joined, either directly, or indirectly via at least one intervening open reading frame, to the 3′ full-length open reading frame to form a composite open reading frame encoding a fusion polypeptide; (b) isolating at least two transformed organisms,exhibiting a different phenotype as compared to a control organism cultivated under the same conditions; (c) isolating at least the 5′ full-length open reading frame and the 3′ open-reading frame from at least two fusion polynucleotides present in the organisms of step (b); (d) randomly combining the isolated full-length open reading frames of step (c) to form at least two random in-frame polynucleotides, each comprising a composite open reading frame encoding a fusion polypeptide; (e) introducing the polynucleotide(s) of step (d) into an organism to produce one or more transformed organisms; and (f) isolating at least one transformed organism from the organism(s) in step (e) exhibiting a different phenotype as compared to a control organism cultivated under the same conditions, wherein steps (c) through (f) are optionally repeated one or more times. |
