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以端粒酶活性为作用靶点筛选抗肿瘤药的一种快速简易方法
A Quickly and Simplified Method for Screening Anticancer Drug in Telomerase Activity as a Target
【摘要】 利用PCR扩增端粒重复序列,通过电泳和银染色测定扩增产物,以电泳胶中6bp梯状条带确定端粒酶活性。温度30℃,反应时间为60min时可以使酶提液与未知化合物作用后还保持着酶的活性。从电泳的上样量及不同浓度细胞酶提液的结果表明,最低上样量10μl和细胞酶提液05μg时,能得到清晰6bp的梯状条带。RNase0.05μg对端粒酶活性显示抑制作用,而对TaqDNA酶没有影响。
【Abstract】 Telomerase activity was measured by the polymerase chain reaction(PCR) based telomeric repeat amplification protocol(TRAP). For analysis of the TRAP assay, the PCR products were separated by the gel electrophoresis. The 6bp DNA ladder was showed by the silver staining. Telomerase activity was observed at the temperature of 30℃ and in reaction time of 60 min. The results obtained varying PCR products quantities and telomerase extract from cell showed that distinct 6bp DNA ladder could be got at 10 μl PCR product quantity and 0.5 μg telomerase extract from tumor cell. Telomerase activity was inhibited at the concentration of 0.05 μg RNase, and no effect occurred for the Taq DNA polymerase.
【Key words】 Telomerase activity; Human tumor cell line; PCR amplification; Screerning durg;
- 【文献出处】 中国药科大学学报 ,JOURNAL OF CHINA PHARMACEUTICAL UNIVERSITY , 编辑部邮箱 ,1999年01期
- 【分类号】R285.5
- 【被引频次】6
- 【下载频次】264