| 摘要 |
Site-specific DNA recombination in eukaryotic cells in vitro comprising using Cre recombinase that lacks heterologous protein-transduction domains, is new. Site-specific DNA recombination in eukaryotic cells in vitro comprises: (a) introducing a polypeptide (I) with Cre recombinase activity but not containing any heterologous protein-transduction domains, into a eukaryotic cell culture where the cells contain DNA region(s) with recognition site(s) for (I); and (b) culturing the cells under suitable conditions for DNA recombination. Independent claims are also included for: (1) a polypeptide (Ia) with Cre recombinase activity comprising fully defined sequences of 343 amino acids (S2) or 371 amino acids (S4), as given in the specification, or their derivatives, homologues and active fragments, excluding the native enzyme from phage P1; (2) nucleic acid sequences (II) comprising fully defined sequences of 1032 base pairs (S1) or 1116 base pairs (S3), as given in the specification, encoding (I); (3) isolated cells that contain (Ia) or (II); (4) a reporter system for detecting site-specific DNA recombination in eukaryotic cells; (5) isolated cells containing the reporter system of (4); (6) preparing (Ia) by recombinant expression; (7) a kit for the new process or the method of (6); and (8) use of glycerol for performing or improving protein transduction.
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