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    • 10. 发明授权
    • Methods and compositions for identifying nucleic acid molecules using nucleolytic activities and hybridization
    • 用核酸分解活性和杂交鉴定核酸分子的方法和组合物
    • US06900013B1
    • 2005-05-31
    • US09648081
    • 2000-08-25
    • Guoqing WangLei WuXiaobo WangJing ChengWeiPing Yang
    • Guoqing WangLei WuXiaobo WangJing ChengWeiPing Yang
    • C07H21/02C07H21/04C12M1/34C12P19/34C12Q1/68
    • C12Q1/6816C07H21/04C12Q1/6827C12Q1/6837C12Q2521/101C12Q2533/101C12Q2563/107C12Q2521/327C12Q2537/143C12Q2565/501
    • The present invention recognizes that identifying genes expressed during developmental processes, stress responses, and disease states can advance understanding of these biological functions, and can contribute to identifying targets for therapeutic drugs. In addition, the present invention recognizes that rapid and reliable profiling of genetic variations, such as mutations and SNPs, is of increasing importance to diagnostics, prognostics, forensics, heredity determinations, and pharmacogenetics. One aspect of the present invention provides a method of identifying one or more nucleic acid molecules that are expressed under a given set of conditions based on their complementarity to known sequences, or one or more mutations or SNPs in a population of nucleic acid molecules. The method includes: contacting at least one probe nucleic acid molecule with a survey population of nucleic acid molecules under conditions that promote nucleic acid hybridization to generate a probe-survey population mixture of nucleic acid molecules, treating the probe-survey population mixture of nucleic acid molecules with a nucleolytic activity, such that nucleolytic activity-sensitive nucleic acid molecules are digested, and contacting the resulting mixture of nucleolytic activity-protected nucleic acid molecules with a solid support comprising one or more attached nucleic acid molecules to generate attached nucleic acid molecule/nucleolytic activity-protected nucleic acid molecule complexes, and identifying one or more of the attached nucleic acid molecules or one or more of the nucleolytic activity-protected nucleic acid molecules in one or more attached nucleic acid molecule/nucleolytic activity-protected nucleic acid molecule complexes.
    • 本发明认识到,在发育过程,应激反应和疾病状态期间表达的鉴定基因可以促进对这些生物功能的理解,并且可以有助于鉴定治疗药物的靶标。 此外,本发明认识到,遗传变异的快速和可靠的分析,例如突变和SNP,对于诊断,预后,取证,遗传测定和药物遗传学是越来越重要的。 本发明的一个方面提供了鉴定一种或多种核酸分子的方法,所述核酸分子基于它们与已知序列的互补性或核酸分子群体中的一个或多个突变或SNP在给定条件下表达。 该方法包括:在促进核酸杂交以产生核酸分子的探针 - 调查群体混合物的条件下使至少一个探针核酸分子与核酸分子的调查群体接触,处理核酸的探针 - 调查群体混合物 具有核分解活性的分子,使得消解活性敏感的核酸核酸分子,并使得到的核分解活性保护的核酸分子的混合物与包含一个或多个连接的核酸分子的固体支持物接触,以产生附着的核酸分子/ 核分解活性保护的核酸分子复合物,以及鉴定一个或多个连接的核酸分子/核酸分解活性保护的核酸分子复合物中所附连的核酸分子或一个或多个核裂解活性保护的核酸分子 。