当前位置: X-MOL 学术Analyst › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Portable glucose meter-utilized label-free and washing-free telomerase assay.
Analyst ( IF 4.2 ) Pub Date : 2020-06-22 , DOI: 10.1039/d0an00655f
Hyo Yong Kim 1 , Chang Yeol Lee , Hansol Kim , Ki Soo Park , Hyun Gyu Park
Affiliation  

We herein describe a portable glucose meter (PGM)-utilized label-free and washing-free method for the facile determination of telomerase activity that relies on the kinase-catalyzed cascade enzymatic reaction (KCER) that transduces the telomerase activity to the glucose level. In the sensor, the telomerase that elongates telomere sequences ((TTAGGG)n) from the 3′-terminus of telomerase substrate primer (TSP) consumes deoxynucleoside triphosphate (dNTP), which serves as a phosphate source for KCER promoted by hexokinase and pyruvate kinase. Thus, the presence of telomerase protects KCER from working effectively, resulting in the maintenance of an initial, high glucose level that is readily determined using hand-held PGM. With this strategy, the telomerase activities in various types of cell lines were successfully determined with high sensitivity. Furthermore, the ability of this method to screen candidate inhibitors for telomerase activity was also verified.

中文翻译:

便携式血糖仪利用的无标记和免洗端粒酶测定法。

我们在本文中描述了一种便携式葡萄糖计(PGM)-利用的无标记和免洗的方法来轻松确定端粒酶活性,该方法依赖于将端粒酶活性转导至葡萄糖水平的激酶催化级联酶促反应(KCER)。在传感器中,延长端粒序列的端粒酶((TTAGGG)n)从端粒酶底物引物(TSP)的3'末端消耗了脱氧核苷三磷酸(dNTP),它是由己糖激酶和丙酮酸激酶促进的KCER的磷酸盐来源。因此,端粒酶的存在使KCER无法有效工作,从而维持了初始的高葡萄糖水平,而使用手持式PGM可以容易地确定该水平。通过这种策略,可以高灵敏度成功地确定各种类型细胞系中的端粒酶活性。此外,还证实了该方法筛选端粒酶活性候选抑制剂的能力。
更新日期:2020-08-10
down
wechat
bug