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    • 6. 发明授权
    • Methods for identifying genomic equivalent markers and their use in quantitating cells and polynucleotide sequences therein
    • 用于鉴定基因组等同标记物的方法及其在定量其中的细胞和多核苷酸序列中的用途
    • US06235504B1
    • 2001-05-22
    • US09481288
    • 2000-01-11
    • Linqi ZhangSharon R. LewinLeondios KostrikisDavid D. Ho
    • Linqi ZhangSharon R. LewinLeondios KostrikisDavid D. Ho
    • C12P1934
    • C12Q1/705C12Q1/6851C12Q1/6883C12Q1/703C12Q2600/156C12Q2600/16
    • Methods for identifying genetic sequences useful as genomic equivalent markers for organisms are described. The method involves determining the ratio of the absolute number of copies of wild type and mutant amplicons in a number of samples from organisms heterozygous for the mutation. After establishing the number of copies of a particular genetic sequence per genome, the sequence may be used as a measure of the number of genomes per sample, in order to normalize the analysis of another target sequence to abundance per cell. By way of example, the CCR5 gene was shown to be present at 2 copies per genome, and used to measure the number of copies per cell of HIV-1 provirions, human herpesvirus-8, and &agr; deletion circles, a measure of recent thymic emigrants for assessing immune reconstitution. The genomic equivalent marker may be use to identify other genomic equivalent markers based on their copy number in proportion to a previously established marker; by way of example the copy number of the &bgr;-actin gene was found to be 16 copies per genome. The genomic equivalent marker may also be use to determine number of cells in a sample, such as from a tissue sample.
    • 描述用于鉴定用作生物体的基因组等效标记物的遗传序列的方法。 该方法包括确定在突变中杂合的生物体的数量的样品中野生型和突变型扩增子的绝对数量的比例。 在确定每个基因组特定遗传序列的拷贝数之后,该序列可以用作每个样品的基因组数量的量度,以便将另一个靶序列的分析归一化为每个细胞丰度。 举例来说,CCR5基因显示以每个基因组2个拷贝存在,并用于测量HIV-1发病细胞,人疱疹病毒-8和α缺失圈的每个细胞的拷贝数,这是最近胸腺的量度 移民用于评估免疫重建。 基因组等价标记可用于基于其与先前建立的标记成比例的拷贝数来鉴定其他基因组等价标记; 通过实例,发现β-肌动蛋白基因的拷贝数为每个基因组16个拷贝。 基因组等同标记物也可用于确定样品中的细胞数,例如来自组织样品。