公开(公告)号：US08114980B2

公开(公告)日：2012-02-14

申请号：US10826522

申请日：2004-04-16

申请人： Geert Plaetinck ,  Christ Platteeuw ,  Katharine Mortier ,  Thierry Bogaert

发明人： Geert Plaetinck ,  Christ Platteeuw ,  Katharine Mortier ,  Thierry Bogaert

IPC分类号： C07H21/04 ,  C12N15/11 ,  G01N33/569 ,  C12Q1/68

CPC分类号： C12N15/1079 ,  C12N15/1093 ,  C12N15/8216 ,  C12N15/8218 ,  C12N15/8279 ,  C12N15/8285 ,  C12N15/86 ,  C12N2795/10243 ,  C12N2800/40 ,  C12N2830/002 ,  C12N2830/008 ,  C12N2830/42 ,  C12Q1/68 ,  Y02A40/164

摘要： There is provided a method of identifying DNA responsible for conferring a particular phenotype in a cell which method comprises a) constructing a cDNA or genomic library of the DNA of the cell in a suitable vector in an orientation relative to a promoter(s) capable of initiating transcription of the cDNA or DNA to double stranded (ds) RNA upon binding of an appropriate transcription factor to the promoter(s), b) introducing the library into one or more of the cells comprising the transcription factor, and c) identifying and isolating a particular phenotype of the cell comprising the library and identifying the DNA or cDNA fragment from the library responsible for conferring the phenotype. Using this technique it is also possible to assign function to a known DNA sequence by a) identifying a homologue(s) of the DNA sequence in a cell, b) isolating the relevant DNA homologue(s) or a fragment thereof from the cell, c) cloning the homologue or fragment thereof into an appropriate vector in an orientation relative to a suitable promoter(s) capable of initiating transcription of dsRNA from the DNA homologue or fragment upon binding of an appropriate transcription factor to the promoter(s) and d) introducing the vector into the cell from step a) comprising the transcription factor.

摘要翻译： 提供了鉴定负责赋予细胞中特定表型的DNA的方法，该方法包括a）在合适的载体中构建相对于能够启动子的启动子的方向的细胞DNA的cDNA或基因组文库 在将合适的转录因子与启动子结合时，启动cDNA或DNA转录为双链（ds）RNA; b）将文库引入至一个或多个包含转录因子的细胞中，c）鉴定和 分离包含文库的细胞的特定表型，并从负责表型的文库鉴定DNA或cDNA片段。 使用这种技术还可以通过以下方式将功能分配给已知的DNA序列：a）鉴定细胞中DNA序列的同系物，b）从细胞中分离相关的DNA同系物或其片段， c）相对于合适的启动子（其能够在合适的转录因子与启动子结合时能够启动dsDNA从DNA同系物或片段的转录）的方向，将其同源物或片段克隆到合适的载体中，并且d ）将载体从包含转录因子的步骤a）引入细胞。