| 摘要 |
Vector (A) for expressing a protein (I) that can activate or inhibit, directly or indirectly, transcription of a reporter gene (RG) present in a host cell comprising: (i) nucleotide sequences encoding a DNA-binding domain (DBD) and a transcription activating domain (TAD), but containing no domains for nuclear localization or nuclear export, and (ii) at least one cloning site for inserting a nucleic acid (X) for screening, is new. Vector (A) for expressing a protein (I) that can activate or inhibit, directly or indirectly, transcription of a reporter gene (RG) present in a host cell comprising: (i) nucleotide sequences encoding a DNA-binding domain (DBD) and a transcription activating domain (TAD), but containing no domains for nuclear localization or nuclear export, and (ii) at least one cloning site for inserting a nucleic acid (X) for screening, where: elements (i) and (ii) are placed under control of a promoter that is inducible and/or repressed by at least one external stimulus so that, after introduction of (X), a protein is expressed that is a fusion of DBD, TAD and the X-encoded product, is new. Independent claims are also included for the following: (a) recombinant plasmid (B) comprising (A) and (X) that is being screened for control of nuclear-cytoplasmic translocation of a given protein, in response to a specific stimulus, with X inserted into the cloning site of the vector; (b) libraries of (B), containing a library of nucleic acids as X; (c) cells transformed with (B) or the library of (b), also including an RG and/or a selection gene (SG), either as a second vector or integrated into the genome; (d) screening nucleotide sequences to identify new proteins or polypeptides for which the nuclear-cytoplasmic distribution is changed in response to a particular stimulus; (e) identifying polypeptide domains and/or amino acids in a particular protein that are involved in control of the nuclear-cytoplasmic distribution in response to a particular stimulus; (f) screening for compounds that block or stimulate the nuclear-cytoplasmic translocation of a particular protein; (g) identifying non-chemical stimuli that block or stimulate the nuclear-cytoplasmic translocation of a particular protein; (h) nucleic acid sequences identified by these methods; (i) kits for performing methods (e)-(g); and (j) kit for detecting or quantifying a stress or toxic agent that includes (A).
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