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    • 6. 发明申请
    • SECRETED AND CELL SURFACE GENES EXPRESSED IN BENIGN AND MALIGNANT COLORECTAL TUMORS
    • 分泌和细胞表面基因表达于胆管和恶性色素性肿瘤
    • WO2003022863A1
    • 2003-03-20
    • PCT/US2002/028518
    • 2002-09-09
    • THE JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINEBUCKHAULTS, PhillipKINZLER, Kenneth, W.VOGELSTEIN, Bert
    • BUCKHAULTS, PhillipKINZLER, Kenneth, W.VOGELSTEIN, Bert
    • C07H21/04
    • C07K14/495C07K14/47C07K14/705
    • Serial analysis of gene expression (SAGE) was used to identify transcripts encoding secreted or cell-surface proteins that were expressed in benign and malignant tumors of the colorectum. A total of 290,394 tags were analyzed from normal, adenomatous and cancerous colonic epithelium. Of the 21,343 different transcripts observed, 957 were found to be differentially expressed between normal and adenoma or between normal and cancer. Forty-nine transcripts were elevated ≥ 20-fold in adenomas, 40 transcripts were elevated ≥ 20-fold in cancers, and nine transcripts were elevated ≥ 20-fold in both. Product of six these nine transcripts (TGFBI, LYS, RDP, MIC-1, REGA, and DEHL) were predicted to be secreted or to reside on the cell surface and these were analyzed in more detail. The abnormal expression levels predicted by SAGE were confirmed by quantitative PCR analyses of each of these six genes. Moreover, the cell types responsible for the elevated expression were identified by in situ hybridization and by PCR analyses of epithelial cells immunoaffinity purified from primary tumors.
    • 基因表达(SAGE)的序列分析用于鉴定在结肠直肠癌和恶性肿瘤中表达的编码分泌的或细胞表面蛋白的转录物。 从正常,腺瘤和癌性结肠上皮分析总共290,394个标签。 在观察到的21,343种不同的转录物中,957被发现在正常和腺瘤之间或在正常和癌症之间差异表达。 在腺瘤中,49个转录本升高了20倍,40个转录物在癌症中升高了20倍,而9个转录物在两者中升高了20倍。 预计这六种转录本(TGFBI,LYS,RDP,MIC-1,REGA和DEHL)的六种产物被分泌或驻留在细胞表面上,并且更详细地分析这些转录物的产物。 通过对这6种基因中的每一种进行定量PCR分析证实SAGE预测的异常表达水平。 此外,通过原位杂交和通过从原发性肿瘤纯化的上皮细胞免疫亲和素的PCR分析来鉴定负责升高的表达的细胞类型。
    • 9. 发明申请
    • MAMMALIAN REGULATOR OF NONSENSE-MEDIATED RNA DECAY
    • 非延迟介导的RNA抑制剂的MAMMALIAN调节剂
    • WO1997040855A1
    • 1997-11-06
    • PCT/US1996015769
    • 1996-10-01
    • THE JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINE
    • THE JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINEDIETZ, Harry, C.
    • A61K48/00
    • G01N33/5008A61K38/00A61K48/00C07K2319/00C12N9/14G01N33/5091G01N33/68
    • All eukaryotes that have been studied to date possess the ability to detect and degrade transcripts that contain a premature signal for the determination of translation. This process of nonsense-mediated RNA decay (NMRD) has been most comprehensively studied in the yeast Saccharomyces cerevisiae where at least three trans-acting factors (Upf1p through Upf3p) are required. The present invention provides cDNAs encoding human and murine RENT1( re gulator of n onsense t ranscripts). RENT1 is the first identified mammalian protein that contains all of the putative functional elements in Upf1p including zinc finger-like motifs and NTPase domains as well as all motifs common to members of helicase superfamily I. Moreover, expression of a chimeric protein, containing the central region of RENT1 flanked by the extreme N- and C-termini of Upf1p, complements the Upf1p-deficient phenotype in yeast.
    • 迄今为止研究的所有真核生物具有检测和降解包含用于确定翻译的过早信号的转录物的能力。 在酵母酿酒酵母(Saccharomyces cerevisiae)中对这种无义介导的RNA衰变(NMRD)的这一过程进行了最全面的研究,其中需要至少三种反式作用因子(Upf1p至Upf3p)。 本发明提供了编码人和鼠RENT1(正义转录本)的cDNA的cDNA。 RENT1是第一种鉴定的哺乳动物蛋白质,其中包含Upf1p中的所有推定的功能元件,包括锌指状基序和NTPase结构域以及解旋酶超家族成员共同的所有基序。此外,含有中枢的嵌合蛋白 RENT1的侧翼是Upf1p的极端N-和C-末端的区域,补充了酵母中的Upf1p缺陷表型。