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1. 发明授权

EP2110440B1 Regulated vectors for controlling DNA hypermutability in eukaryotic cells 有权
标题翻译：用于DNA超突变的真核细胞中控制调节的矢量
公开(公告)号：EP2110440B1
公开(公告)日：2010-12-22
申请号：EP09155285.1
申请日：2003-02-21
申请人： Morphotek, Inc.
发明人： Grasso, Luigi , Nicholaides, Nicholas C , Sass, Philip M
IPC分类号： C12Q1/68 , C12N15/85
CPC分类号： C12N15/1024 , C12N15/85 , C12N2830/42 , C12N2840/203

2. 发明公开

EP2110440A3 Regulated vectors for controlling DNA hypermutability in eukaryotic cells 有权
标题翻译：用于DNA超突变的真核细胞中控制调节的矢量
公开(公告)号：EP2110440A3
公开(公告)日：2009-10-28
申请号：EP09155285.1
申请日：2003-02-21
申请人： Morphotek, Inc.
发明人： Grasso, Luigi , Nicholaides, Nicholas C , Sass, Philip M
IPC分类号： C12Q1/68 , C12N15/85
CPC分类号： C12N15/1024 , C12N15/85 , C12N2830/42 , C12N2840/203
摘要： 1. The present invention provides a method for producing an isolated, genetically stable somatic cell line with a new phenotype, the method comprising the steps of:
a) culturing a recombinant host somatic cell comprising a vector, said vector comprising a constitutively active promoter operatively linked to a sequence encoding a dominant negative allele of a mismatch repair gene, an internal ribosome entry site, and a negative selection marker, under conditions for the expression of the dominant negative allele, rendering the recombinant host somatic cell hypermutable and thereby producing a library of cells;
b) selecting for clones from the cell library exhibiting new phenotypes;
c) from the clones selected in step (b), negatively selecting for clones no longer expressing the dominant negative allele of a mismatch repair gene, rendering the resulting subclones genetically stable; and
d) expanding a genetically stable subclone of step (c) to generate said genetically stable cell line.

3. 发明公开

EP2110440A2 Regulated vectors for controlling DNA hypermutability in eukaryotic cells 有权
标题翻译：用于控制真核细胞中DNA超可变性的调控载体
公开(公告)号：EP2110440A2
公开(公告)日：2009-10-21
申请号：EP09155285.1
申请日：2003-02-21
申请人： Morphotek, Inc.
发明人： Grasso, Luigi , Nicholaides, Nicholas C , Sass, Philip M
IPC分类号： C12Q1/68 , C12N15/85
CPC分类号： C12N15/1024 , C12N15/85 , C12N2830/42 , C12N2840/203
摘要： 1. The present invention provides a method for producing an isolated, genetically stable somatic cell line with a new phenotype, the method comprising the steps of:
a) culturing a recombinant host somatic cell comprising a vector, said vector comprising a constitutively active promoter operatively linked to a sequence encoding a dominant negative allele of a mismatch repair gene, an internal ribosome entry site, and a negative selection marker, under conditions for the expression of the dominant negative allele, rendering the recombinant host somatic cell hypermutable and thereby producing a library of cells;
b) selecting for clones from the cell library exhibiting new phenotypes;
c) from the clones selected in step (b), negatively selecting for clones no longer expressing the dominant negative allele of a mismatch repair gene, rendering the resulting subclones genetically stable; and
d) expanding a genetically stable subclone of step (c) to generate said genetically stable cell line.
摘要翻译： 1.本发明提供了产生具有新表型的分离的，遗传稳定的体细胞系的方法，所述方法包括以下步骤：a）培养包含载体的重组宿主体细胞，所述载体在操作上包含组成型活性启动子在显性失活等位基因表达的条件下，与编码错配修复基因的显性失活等位基因，内部核糖体进入位点和阴性选择标记的序列相连，使重组宿主体细胞超可变，从而产生文库细胞; b）从展示新表型的细胞文库中选择克隆; c）从步骤（b）中选择的克隆中，阴性选择不再表达错配修复基因的显性失活等位基因的克隆，使得到的亚克隆遗传稳定; 和d）扩增步骤（c）的遗传稳定的亚克隆以产生所述遗传稳定的细胞系。

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