水稻端粒酶活性定量分析研究

摘要：端粒酶活性直接与细胞的分生能力、生活力、生命力有着密不可分的关系， 因此对其活性测定有着重要意义。本文参照人类端粒酶体外检测的原理和方法，设计了特别的先导引物和反向引物，采用不同的退火温度将PCR循环分成两步进行，再结合DNA凝胶电泳成像定量分析系统，以模式植物水稻为研究对象，对端粒酶活性定量检测方法和反应条件进行了探索。结果显示，水稻端粒酶活性的最佳反应条件为：温度19℃，反应时间13分钟，总蛋白浓度0.28μg/μl，与人端粒酶最佳反应条件有明显差异，建立了一种有效测定植物端粒酶活性的定量检测方法。应用该方法对6个水稻品种的根、幼叶及幼穗三个不同组织器官的端粒酶活性进行了定量测定，结果显示幼穗最高，其次为幼叶，根最弱。说明植物端粒酶活性与细胞、组织的生活力有着密切关系。

Quantitative Analysis for Telomerase Activtity in Rice

The Telomeric Repeat Amplification Protocol (TRAP) and its modified versions change the size and/or the ratio of the telomerase products in the amplification stage of the assay. Based on TRAP, we developed a useful method for detecting telomerase activity in rice. In this report we designed a special precursor primer and a special reverse primer and conducted two steps of PCR cycles. GENE Genius? Bio-imaging System was applied for this quantitative analysis for exploring telomerase activity and its optimal reaction conditions. The method ensured that the optimal reaction conditions for the telomerase was 19℃,13minutes, at a concentration of 0.28 u g/μ l. A quantitative analysis method was established for detecting telomerase activity in rice. With this method, we detected telomerase activity in roots, young leaves and young panicles of six parental lines of hybrid rice. The results show that young panicles have the highest telomerase activity, demonstrating that telomerase activity is closely related to the cell vitality in plants.