公开(公告)号：US20050037351A1

公开(公告)日：2005-02-17

申请号：US10483071

申请日：2002-07-09

申请人： Kimiyoshi Kanno ,  Naozumi Oda ,  Masaharu Aritomi ,  Akiko Sato

发明人： Kimiyoshi Kanno ,  Naozumi Oda ,  Masaharu Aritomi ,  Akiko Sato

IPC分类号： C12N15/10 ,  C12Q1/68 ,  C07H21/04

CPC分类号： C12Q1/6806 ,  C12N15/1017 ,  C12Q2523/308

摘要： A method of separating and purifying nucleic acids from samples containing cells, such as blood and culture solutions. According to the method of the invention, a cell extract obtained by cell disruption is adsorbed by a filter made of a nonwoven fabric and the nucleic acid is eluted after washing the filter. Alkaline conditions of pH 12 or higher may be employed for elution of the nucleic acid, or the filter-adsorbed nucleic acid may be eluted by treatment with active oxygen or by using a surfactant. Nucleic acids separated and purified by the method of the invention can be used in nucleic acid amplification and nucleic acid sequence analysis techniques.

摘要翻译： 从含有细胞例如血液和培养液的样品中分离和纯化核酸的方法。 根据本发明的方法，通过细胞破碎获得的细胞提取物被无纺布制成的过滤器吸附，并且在洗涤过滤器之后洗脱核酸。 可以使用pH12或更高的碱性条件来洗脱核酸，或者通过用活性氧处理或通过使用表面活性剂洗脱滤器吸附的核酸。 通过本发明方法分离和纯化的核酸可用于核酸扩增和核酸序列分析技术。

公开(公告)号：US06214613B1

公开(公告)日：2001-04-10

申请号：US08647924

申请日：1996-07-12

申请人： Kazuo Higuchi ,  Kimiyoshi Kanno

发明人： Kazuo Higuchi ,  Kimiyoshi Kanno

IPC分类号： C12N1500

CPC分类号： C40B40/02 ,  C07K16/00 ,  C07K16/1282 ,  C07K16/4283 ,  C12N15/1037 ,  C12N15/85 ,  C12N2800/108

摘要： An expression vector for preparing a library of an antibody variable region, which can express comprising polypeptides containing the H-chain and/or L-chain variable regions of antibodies in the membrane-bound form on the surfaces of eukaryotic cells, which is containing the nucleotide sequence of AKL (formula 1) and/or the nucleotide sequence AKH (formula 2), and which is replicable in the cells: 5′-PL-SL-CL-ML-AL-3′  (formula 1)(AKL) 5′-PH-SH-CH-MH-AH-3′  (formula 2)(AKH) wherein PL and PH represent a promoter, respectively; SL and SH represent a nucleotide sequence coding for a signal peptide respectively; CL represents a nucleotide sequence coding for the L-chain constant region of an antibody; CH represents a nucleotide sequence coding for the H-chain constant region of an antibody or a nucleotide sequence coding for a polypeptide containing at least CH1 of the H-chain constant region of an antibody; AL and AH represent a polyadenylation signal, respectively; “5′-” and “-3′” refer to the 5′ side and the 3′ side, respectively, of the nucleotide sequence; and ML and MH represent a nucleotide sequence coding for the transmembrane domain, respectively, while either ML or MH may be a mere chemical bond when the expression vector contains the nucleotide sequence of AKL and AKH. Cloning site of R1L, R2L, R1H and R2H is present within or in the vicinity of SL, CL, SH and CH respectively, in order for the nucleotide sequences coding for the L-chain and H-chain variable regions to be readily inserted in between SL and CL, and SH and CH respectively.

摘要翻译： 用于制备抗体可变区的文库的表达载体，其可以表达包含含有在真核细胞表面上的膜结合形式的抗体的H链和/或L链可变区的多肽，其含有 AKL（式1）和/或核苷酸序列AKH（式2）的核苷酸序列，其可在细胞中复制：其中PL和PH分别代表启动子; SL和SH分别表示编码信号肽的核苷酸序列; CL表示编码抗体的L链恒定区的核苷酸序列; CH表示编码抗体的H链恒定区的核苷酸序列或编码抗体的H链恒定区至少CH1的多肽的核苷酸序列; AL和AH分别表示聚腺苷酸化信号; “5'-”和“-3”分别指核苷酸序列的5'侧和3'侧; ML和MH分别表示编码跨膜结构域的核苷酸序列，而当表达载体含有AKL和AKH的核苷酸序列时，ML或MH可能仅仅是化学键。 R1L，R2L，R1H和R2H的克隆位点分别存在于SL，CL，SH和CH 3附近或其附近，以便将编码L链和H链可变区的核苷酸序列容易地插入 在SL和CL之间，SH和CH分别。

公开(公告)号：US06777204B1

公开(公告)日：2004-08-17

申请号：US09786442

申请日：2001-03-05

申请人： Tsuneo Takahashi ,  Mitsuharu Ono ,  Hiroshi Ishimaru ,  Kimiyoshi Kanno ,  Chiaki Takahashi

发明人： Tsuneo Takahashi ,  Mitsuharu Ono ,  Hiroshi Ishimaru ,  Kimiyoshi Kanno ,  Chiaki Takahashi

IPC分类号： C12P2106

CPC分类号： C07K14/705 ,  C07K16/28 ,  C07K2317/34

摘要： Disclosed are a novel seven-pass transmembrane receptor protein found in immature dendritic cells and a DNA encoding the same. Further, disclosed are a replicable recombinant DNA which comprises a replicable expression vector and, operably inserted therein, the above-mentioned DNA; a cell of a microorganism or cell culture (transformant), which is transformed with the above-mentioned replicable recombinant DNA; a seven-pass transmembrane receptor protein which is produced on the cell surface of the above-mentioned transformant; a method for screening a ligand which binds to the above-mentioned seven-pass transmembrane receptor protein, and a method for screening a substance which inhibits the ligand from binding to the seven-pass transmembrane receptor protein; and an antibody which binds to the above-mentioned seven-pass transmembrane receptor protein. The present invention also discloses a method for the diagnosis of an inflammatory disease, such as rheumatism, which comprises determining the amount of the seven-pass transmembrane receptor protein expressed in human leukocytes.

摘要翻译： 公开了一种在未成熟树突状细胞中发现的新型七遍跨膜受体蛋白和编码它的DNA。 此外，公开了可复制的重组DNA，其包含可复制的表达载体，并在其中可操作地插入上述DNA; 用上述可复制重组DNA转化的微生物或细胞培养物（转化体）的细胞; 在上述转化体的细胞表面产生的7次跨膜受体蛋白; 用于筛选结合上述七次跨膜受体蛋白质的配体的方法，以及筛选抑制配体与七次跨膜受体蛋白结合的物质的方法; 和结合上述七次跨膜受体蛋白的抗体。 本发明还公开了一种用于诊断炎性疾病如风湿病的方法，其包括测定人白细胞中表达的七次跨膜受体蛋白的量。