One-step affinity purification protocol for human telomerase

被引：76

作者：

Schnapp, G ^{[1
]}

Rodi, HP ^{[1
]}

Rettig, WJ ^{[1
]}

Schnapp, A ^{[1
]}

Damm, K ^{[1
]}

机构：

[1] Boehringer Ingelheim Pharma KG, Dept Oncol Res, D-88397 Biberach, Germany

来源：

NUCLEIC ACIDS RESEARCH | 1998年 / 26卷 / 13期

关键词：

D O I：

10.1093/nar/26.13.3311

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Human telomerase is a ribonucleoprotein (RNP) enzyme, comprising protein components and an RNA template that catalyses telomere elongation through the addition of TTAGGG repeats. Telomerase function has been implicated in aging and cancer cell immortalization. We report a rapid and efficient one-step purification protocol to obtain highly active telomerase from human cells. The purification is based on affinity chromatography of nuclear extracts with antisense oligonucleotides complementary to the template region of the human telomerase RNA component. Bound telomerase is eluted with a displacement oligonucleotide under mild conditions. The:resulting affinity-purified telomerase is active in PCR-amplified telomerase assays. The purified telomerase complex has a molecular mass of similar to 550 kDa compared to the similar to 1000 kDa determined for the telomerase RNP in unfractionated nuclear extracts. The purification protocol provides a rapid and efficient tool for functional and structural studies of human telomerase.

引用

页码：3311 / 3313

页数：3

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