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    • 6. 发明授权
    • Antioxidant responsive element
    • 抗氧化反应元件
    • US6120994A
    • 2000-09-19
    • US862431
    • 1997-05-23
    • Shui-Pang Tam
    • Shui-Pang Tam
    • C12Q1/68C07H21/04
    • C12Q1/6897
    • The present invention relates, in general, to antioxidant responsive elements (AREs). In particular, the present invention relates to a DNA construct comprising an ARE having the DNA sequence 5'-RGR AC NNN GCT-3' (SEQ ID NO: 1) operably linked to a heterologous protein coding sequence; cells and non-human organisms comprising the DNA construct; a method of screening for a compound that increases transcription of an MRNA regulated by an antioxidant responsive element; and a purified compound that binds to an antioxidant responsive element.
    • 本发明一般涉及抗氧化反应元件(ARE)。 特别地,本发明涉及包含与异源蛋白质编码序列可操作地连接的具有DNA序列5'-RGRACNNN GCT-3'(SEQ ID NO:1)的ARE的DNA构建体。 包含DNA构建体的细胞和非人生物; 筛选增加由抗氧化剂反应元件调节的MRNA的转录的化合物的方法; 和与抗氧化反应元件结合的纯化化合物。
    • 7. 发明授权
    • DNA constructs and methods for screening for increased expression of
human apo AI gene
    • 用于筛选人apo AI基因表达增加的DNA构建体和方法
    • US5994061A
    • 1999-11-30
    • US536559
    • 1995-09-29
    • Shui-Pang TamXia Zhang
    • Shui-Pang TamXia Zhang
    • C12Q1/68
    • C12Q1/6897
    • Methods for screening for a drug that increases expression of the human apolioprotein (apo) AI gene and related DNA constructs. A first method has the steps of: (a) introducing into mammalian cells a DNA construct including, functionally joined together in the 5'.fwdarw.3' direction of transcription, (i) at least one copy of a drug-responsive element (DRE) having at least about 80% homology with the DNA sequence 5'-G/C N T/G A/G GCTGGG-3', (ii) a heterologous promoter, (iii) a reporter gene and (iv) an untranslated region including a functional polyadenylation signal; (b) growing a first culture of the cells in the absence of drug; (c) lysing the first culture to produce a first extract; (d) assaying the first extract for activity of a protein encoded by the reporter gene; (e) growing a second culture of the cells in the presence of the drug; (f) lysing the second culture to produce a second extract; (g) assaying the second extract for activity of the protein encoded by the reporter gene; and (h) comparing the activities of the first extract and the second extract. A second method uses a different DNA construct including (i) a promoter region of the human apo AI gene including at least one copy of a drug-responsive element (DRE) having at least about 80% homology with the DNA sequence 5'-G/C N T/G A/G GCTGGG-3', (ii) a reporter gene and (iii) an untranslated region including a functional polyadenylation signal. A third method involves stably maintaining the introduced DNA in the cells.
    • 筛选增加人apolioprotein(apo)AI基因和相关DNA构建体表达的药物的方法。 第一种方法具有以下步骤:(a)向哺乳动物细胞中引入包括在转录的5''3'方向上功能连接在一起的DNA构建体,(i)至少一个药物反应元件(DRE )具有与DNA序列5'-G / CNT / GA / G GCTGGG-3'至少约80%的同源性,(ii)异源启动子,(iii)报告基因和(iv)非翻译区, 多聚腺苷酸化信号; (b)在没有药物的情况下培养细胞的第一培养物; (c)裂解第一培养物以产生第一提取物; (d)测定第一提取物对报道基因编码的蛋白质的活性; (e)在药物存在下培养细胞的第二培养物; (f)裂解第二培养物以产生第二提取物; (g)测定第二提取物对报道基因编码的蛋白质的活性; 和(h)比较第一提取物和第二提取物的活性。 第二种方法使用不同的DNA构建体,其包括(i)人apo AI基因的启动子区域,包括与DNA序列5'-G具有至少约80%同源性的药物应答元件(DRE)的至少一个拷贝 / CNT / GA / G GCTGGG-3',(ii)报告基因和(iii)包含功能多聚腺苷酸化信号的非翻译区。 第三种方法包括稳定地将引入的DNA保持在细胞中。