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1. 发明申请

US20050191666A1 Method of identification and cloning differentially expressed messenger RNAs 审中-公开
标题翻译：识别和克隆差异表达信使RNA的方法
公开(公告)号：US20050191666A1
公开(公告)日：2005-09-01
申请号：US11031077
申请日：2005-01-10
申请人： Alexander Belyavsky , Natalia Ivanova
发明人： Alexander Belyavsky , Natalia Ivanova
IPC分类号： C07H21/02 , C07H21/04 , C12N15/10 , C12P19/34 , C12Q1/68 , C12Q1/6809 , C12Q1/6855
CPC分类号： C12N15/1072 , C12N15/1096 , C12Q1/6809 , C12Q1/6855
摘要： A method of identification of differentially expressed messenger RNA (mRNA) which consists of synthesizing from a set of sequences of mRNA sets of fragments of complementary DNA (cDNA), which are separated with the aid of gel electrophoresis and the pictures of separation of the cDNA from different types of cells are compared and fragments with differential signal intensity are identified. For formation of the set of fragments the cDNA is cleaved with the aid of restriction nucleases. A method of cloning of differentially expressed mRNAs consists of synthesizing from sets of sequences of mRNAs from different types of cells sets of fragments of complementary DNA (cDNA) which are separated with the aid of gel electrophoresis, the pictures of the separation of the cDNA from different types of cells are compared, fragments of cDNA with different signal intensities are separated from the gel, amplified with the aid of a polymerase chain reaction and cloned to a plasmid or phage vector. For the formation of the set of fragments one carried out cleavage of the cDNA with the aid of restriction endonucleases and uses only those fragments of cDNA that correspond to the 3′ or 5′ and regions of the mRNAs.
摘要翻译：鉴别差异表达的信使RNA（mRNA）的方法，其由通过凝胶电泳分离的互补DNA（cDNA）片段的mRNA集合的一组序列合成而组合，以及分离cDNA的图片比较来自不同类型的细胞，并鉴定具有差异信号强度的片段。为了形成该组片段，借助于限制性核酸酶将cDNA切割。克隆差异表达的mRNA的方法包括从借助于凝胶电泳分离的来自不同类型的细胞组的互补DNA片段（cDNA）的mRNA集合的集合的合成，将cDNA从比较不同类型的细胞，从凝胶中分离具有不同信号强度的cDNA片段，借助于聚合酶链反应扩增，并克隆到质粒或噬菌体载体中。为了形成一组片段，一个在限制性内切核酸酶的帮助下进行cDNA的切割，并且仅使用对应于mRNA的3'或5'和区域的那些cDNA片段。

2. 发明授权

US06670123B1 Method for detecting hematopoietic stem cells 失效
标题翻译：检测造血干细胞的方法
公开(公告)号：US06670123B1
公开(公告)日：2003-12-30
申请号：US09706594
申请日：2000-11-03
申请人： Alexander Belyavsky , Sergey Shmelkov , Jan Visser
发明人： Alexander Belyavsky , Sergey Shmelkov , Jan Visser
IPC分类号： C07H2104
CPC分类号： C07K14/70596 , Y10S530/827
摘要： The present invention provides an isolated nucleic acid encoding KIAA0918, an isolated nucleic acid that hybridizes under high stringency conditions to a nucleic acid that is complementary to a nucleic acid encoding KIAA0918, a purified KIAA0918 protein, a purified protein encoded by a nucleic acid that hybridizes under high stringency conditions to a nucleic acid that is complementary to a nucleic acid encoding KIAA0918, a method of making KIAA0918 protein, an antibody specific for KIAA0918, a method for producing an antibody specific for KIAA0918 protein, a vector comprising a nucleic acid encoding KIAA0918, and a host cell transformed with a vector comprising a nucleic acid encoding KIAA0918. Also provided are methods for detecting the presence of and isolating hematopoietic stem cells in a heterogeneous cell suspension and for assessing gene expression in a tissue sample.
摘要翻译：本发明提供了编码KIAA0918的分离的核酸，该分离的核酸在高严格条件下与与编码KIAA0918的核酸互补的核酸，纯化的KIAA0918蛋白，由杂交的核酸编码的纯化蛋白质杂交在高严格条件下，与编码KIAA0918的核酸互补的核酸，制备KIAA0918蛋白质的方法，KIAA0918特有的抗体，KIAA0918蛋白质特异性抗体的制备方法，包含编码KIAA0918的核酸的载体以及用包含编码KIAA0918的核酸的载体转化的宿主细胞。还提供了用于检测异源细胞悬液中的造血干细胞的存在和分离并用于评估组织样品中的基因表达的方法。

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