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[端粒长度和端粒酶活性的测量方法——实践与问题]

[Methods of measuring telomere length and telomerase activity--practice and problems].

作者信息

Saito Y, Suda T, Hatakeyama K

机构信息

First Department of Surgery, Niigata University School of Medicine.

出版信息

Nihon Rinsho. 1998 May;56(5):1153-8.

PMID:9613113
Abstract

The development of a highly sensitive method for detection of telomerase activity, telomeric repeat amplification protocol (TRAP), has provided knowledge on telomerase activity in normal and cancer tissues. Subsequent several modifications have been achieved, including an introduction of the internal standard and hybridization protection technique that leads to simplicity and improvement of reproducibility and linearity of this method, and application of TRAP to in situ analysis to identify the cells responsible for telomerase activity. As for measurement of telomere length, fluorescence in situ hybridization technique appeared to give an information of telomere length on an individual chromosome in contrast to analysis of terminal restriction fragment, a conventional method which can express mean telomere length of all chromosomes. Further methodological improvement in this field is ongoing and showing a new sight on cell mortality and immortality.

摘要

一种用于检测端粒酶活性的高灵敏度方法——端粒重复序列扩增法(TRAP)的发展,为了解正常组织和癌组织中的端粒酶活性提供了依据。随后又进行了多项改进,包括引入内标和杂交保护技术,这使得该方法更加简便,重现性和线性得到提高,并且将TRAP应用于原位分析以鉴定产生端粒酶活性的细胞。至于端粒长度的测量,荧光原位杂交技术似乎能够提供单个染色体上端粒长度的信息,这与传统的分析末端限制片段的方法不同,后者只能表达所有染色体的平均端粒长度。该领域的方法学仍在不断改进,为细胞死亡和永生带来了新的见解。

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