选择单链DNA分子,可以结合和抑制人类血栓激素
L C Bock1, L C Griffin, J A Latham
1Gilead Sciences Inc., Foster City, California 94404.
Nature
|February 6, 1992
概括
研究人员开发了针对血凝中的关键蛋白质 - - 血栓素的单链DNA吸收体. 这些DNA体表现出显著的结合亲和力,并抑制了血栓激素.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 胺是核酸分子,对分子标具有很高的特异性.
- 之前的阿普坦体开发主要利用双链DNA或单链RNA.
- 单链DNA吸附体和与核酸在生理上不相互作用的点在很大程度上仍未被探索.
研究的目的:
- 为了研究单链DNA (ssDNA) 作为体的潜力.
- 为了分离和描述针对血栓素的ssDNA吸收体,这是凝血级联中的关键蛋白酶.
- 为了评估这些阿巴的功能抑制能力对血栓激素活性.
主要方法:
- 实验室选择和聚合酶连锁反应 (PCR) 用于从随机DNA池中丰富aptamer候选物.
- 隔离的阿普坦酶的测序确定了对标结合至关重要的保护区域.
- 使用纳米分子 (nM) 度范围量化结合亲和力.
- 试验室试验评估了血催化纤维素凝块形成的抑制.
主要成果:
- 成功分离了针对蛋白血的单链DNA体.
- 在25-200 nM的范围内获得了对血栓胺体的结合亲和力.
- 在32个被测序的血栓激素体中确定了一个保存的14-17基区域.
- 证明了几种ssDNA体在净化和血样本中的纳米分子度上抑制了血的纤维素凝固活性.
结论:
- 单链DNA可以作为针对蛋白质标的有效体起作用.
- 已识别的ssDNA体表现出高亲和力和功能抑制活性对血栓.
- 这些发现扩大了对生物系统中的蛋白质进行向的aptamer技术的范围,特别是在凝血研究中.
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