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    DNA encoding GM-CSF and a method of producing GM-CSF protein
    7.
    发明授权
    DNA encoding GM-CSF and a method of producing GM-CSF protein 失效
    编码GM-CSF的DNA和生产GM-CSF蛋白的方法

    公开(公告)号：US5908763A

    公开(公告)日：1999-06-01

    申请号：US287019

    申请日：1994-08-08

    申请人： Steven C. Clark ,  Randal J. Kaufman ,  Gordon G. Wong ,  Elizabeth A. Wang

    发明人： Steven C. Clark ,  Randal J. Kaufman ,  Gordon G. Wong ,  Elizabeth A. Wang

    IPC分类号： A61K38/00 ,  C07K14/535 ,  C12N15/27 ,  C12N15/70 ,  C12N15/81 ,  C12N15/85 ,  C07K14/53 ,  C12N15/63

    CPC分类号： C12N15/85 ,  C07K14/535 ,  C12N15/70 ,  C12N15/81 ,  A61K38/00 ,  C12N2800/206 ,  C12N2830/00 ,  C12N2830/702 ,  C12N2840/105 ,  C12N2840/44

    摘要： A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises:preparing RNA from a cell that produces CSF;preparing polyadenylated messenger RNA from said RNA;preparing single stranded cDNA from said messenger RNA;converting the single stranded cDNA to double stranded cDNA;inserting the double stranded cDNA into transformation vectors and transforming bacteria with said vector to form colonies;picking pools of 200 to 500 colonies each and isolating plasmid DNA from each pool;transfecting the plasmid DNA into suitable host cells for expressing CSF protein;culturing the transfected cells and assaying the supernatant for CSF activity; andselecting CSF positive pools and screening the colonies used to make the pool to identify a colony having CSF activity. Also described are a cDNA coding for a protein having CSF activity (i.e. CSF/cDNA), a microorganism or cell line transformed with a recombinant vector containing such CSF/cDNA, and a method for producing CSF protein by expressing said CSF/cDNA by culturing a microorganism or cell line. The invention also provides a method of purifying the CSF proteins and the purified proteins so produced.

    摘要翻译： 描述了制备和分离含有CSF / cDNA的转化载体的方法。 该方法包括：从产生CSF的细胞制备RNA; 从所述RNA制备聚腺苷酸化的信使RNA; 从所述信使RNA制备单链cDNA; 将单链cDNA转化为双链cDNA; 将双链cDNA插入转化载体并用所述载体转化细菌以形成菌落; 分别取200至500个菌落的池，每个池中分离出质粒DNA; 将质粒DNA转染到合适的宿主细胞中以表达CSF蛋白; 培养转染的细胞并测定上清液的CSF活性; 并选择CSF阳性池并筛选用于制备池的菌落以鉴定具有CSF活性的菌落。 还描述了编码具有CSF活性的蛋白质（即CSF / cDNA）的cDNA，用含有这种CSF / cDNA的重组载体转化的微生物或细胞系，以及通过培养表达所述CSF / cDNA来产生CSF蛋白的方法 微生物或细胞系。 本发明还提供了纯化CSF蛋白质和如此制备的纯化蛋白质的方法。

    Expression monitoring by hybridization to high density nucleic acid arrays
    10.
    发明授权
    Expression monitoring by hybridization to high density nucleic acid arrays 有权
    通过与高密度核酸阵列杂交进行表达监测

    公开(公告)号：US06410229B1

    公开(公告)日：2002-06-25

    申请号：US09212004

    申请日：1998-12-14

    申请人： David J. Lockhart ,  Eugene L. Brown ,  Gordon G. Wong ,  Mark S. Chee ,  Thomas R. Gingeras

    发明人： David J. Lockhart ,  Eugene L. Brown ,  Gordon G. Wong ,  Mark S. Chee ,  Thomas R. Gingeras

    IPC分类号： C12Q168

    CPC分类号： C12Q1/6809 ,  C12Q1/6837 ,  G01N15/1475 ,  C12Q2565/507

    摘要： This invention provides methods of monitoring the expression levels of a multiplicity of genes. The methods involve hybridizing a nucleic acid sample to a high density array of oligonucleotide probes where the high density array contains oligonucleotide probes complementary to subsequences of target nucleic acids in the nucleic acid sample. In one embodiment, the method involves providing a pool of target nucleic acids comprising RNA transcripts of one or more target genes, or nucleic acids derived from the RNA transcripts, hybridizing said pool of nucleic acids to an array of oligonucleotide probes immobilized on surface, where the array comprising more than 100 different oligonucleotides and each different oligonucleotide is localized in a predetermined region of the surface, the density of the different oligonucleotides is greater than about 60 different oligonucleotides per 1 cm2, and the olignucleotide probes are complementary to the RNA transcripts or nucleic acids derived from the RNA transcripts; and quantifying the hybridized nucleic acids in the array.

    摘要翻译： 本发明提供监测多种基因的表达水平的方法。 所述方法包括将核酸样品与寡核苷酸探针的高密度阵列杂交，其中高密度阵列含有与核酸样品中靶核酸的子序列互补的寡核苷酸探针。 在一个实施方案中，所述方法包括提供包含一个或多个靶基因的RNA转录物或衍生自RNA转录物的核酸的靶核酸库，将所述核酸库与固定在表面上的寡核苷酸探针阵列杂交，其中 包含超过100种不同寡核苷酸的阵列和每种不同的寡核苷酸被定位在表面的预定区域中，不同寡核苷酸的密度大于每1cm 2大约60个不同的寡核苷酸，并且寡核苷酸探针与RNA转录物互补 衍生自RNA转录物的核酸; 并定量阵列中的杂交核酸。