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    • 4. 发明申请
    • REGULATED STOP CODON READTHROUGH
    • 调节停止CODON READTHROUGH
    • WO2005073375A1
    • 2005-08-11
    • PCT/DK2005/000070
    • 2005-01-28
    • MAXYGEN HOLDINGS LTD.MAXYGEN APSBOUQUIN, Thomas
    • BOUQUIN, Thomas
    • C12N15/10
    • C12N15/1086C07K2319/035C07K2319/40C07K2319/60C12N15/62C12N15/67G01N33/502
    • Methods for screening or selecting cells expressing a polypeptide of interest, e.g. for selecting cells expressing a desired level of a polypeptidé of interest, for evaluating recombinant polypeptide expression in a population of cells, or for selecting cells expressing a polypeptide with a desired binding affinity to a ligand, as well as for producing a polypeptide of interest from a selected cell, where the cells comprise an expression cassette comprising a first polynucleotide encoding the polypeptide of interest, at least one stop codon downstream of the first polynucleotide, and a second polynucleotide encoding a cell membrane anchoring peptide, a reporter peptide or an epitope tag downstream of the stop codon, and where the cells are cultured in the presence of a termination suppression agent, in particular an aminoglycoside antibiotic, that results in regulated stop codon readthrough.
    • 用于筛选或选择表达目的多肽的细胞的方法,例如 用于选择表达期望水平的感兴趣多肽的细胞,用于评估细胞群体中的重组多肽表达,或用于选择表达具有对配体的所需结合亲和力的多肽的细胞,以及用于产生感兴趣的多肽 选择的细胞,其中细胞包含表达盒,其包含编码目标多肽的第一多核苷酸,第一多核苷酸下游的至少一个终止密码子,以及编码细胞膜锚定肽,报告肽或表位标签的第二多核苷酸 在终止密码子的下游,并且其中细胞在终止抑制剂(特别是氨基糖苷类抗生素)存在下培养,导致调节的终止密码子读取。