Abstract
In 1985, telomerase activity was identified in the macronuclei of the ciliate Tetrahymena and was found to add telomeric repeats onto telomeric oligonucleotide primers (1). The radiolabeled elongated products were detected easily as a 6-bp ladder on an electrophoresis gel. In 1989, using the same method, human telomerase activity was identified in a HeLa cell extract (2). However, the telomerase activity of human cells was very weak, and it was difficult to analyze the biochemical properties of telomerase or to investigate the significance of telomerase in various diseases, such as cancer. Therefore, we began to investigate a sensitive polymerase chain reaction (PCR)-based detection method for telomerase activity. Our goal was to develop a method that was not only versatile but also useful for biochemical analyzes, such as the measurement of the level of processing and the kinetics of telomerase.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Greider, C. W. and Blackburn, E. H. (1985) Identification of a specific telomere terminal transferase activity in Tetrahymena extracts. Cell 43, 405–413.
Morin, G. B. (1989) The human telomere terminal transferase enzyme is a ribonucleoprotein that synthesizes TTAGGG repeats. Cell 59, 521–529.
Kim, N. W., Piatyszek, M. A., Prowse, K. R., et al. (1994). Specific association of human telomerase activity with immortal cells and cancer. Science 266, 2011–2015.
Prowse, K. R. and Greider, C. W. (1995). Developmental and tissuespecific regulation of mouse telomerase and telomere length. Proc. Natl. Acad. Sci. USA 92, 4818–4822.
Counter, C. M., Gupta, J., Harley, C. B., Leber, B., and Bacchetti, S. (1995) Telomerase activity in normal leucocytes and in haematologic malignancies. Blood 85, 2315–2320.
Broccoli, D., Young, J. W., and de Lange, T. (1995) Telomerase activity in normal and malignant hematopoietic cells. Proc. Natl. Acad. Sci. USA 92, 9082–9086.
Tatematsu, K., Nakayama, J., Danbara, M., Shionoya, S., Sato, H., Omine, M., and Ishikawa, F. (1996) A novel quantitative’ stretch PCR assay’ that detects a dramatic increase in telomerase activity during the progression of myloid leukemias. Oncogene 13, 2265–2274.
Nakayama, J., Saito, M., Nakamura, H., Matsuura, A., and Ishikawa, F. (1997) TLP1: a gene encoding a protein component of mammalian reverse telomerase is a novel member of WD repeat family. Cell 88, 875–884.
Nakayama, J., Tahara, H., Tahara, E., Saito, M., et al. (1998) Telomerase activation by hTRT in normal human fibroblasts and hepatocellular carcinomas. Nat. Genet. 18, 65–68.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Nakayama, Ji., Ishikawa, F. (2002). Stretch PCR Assay. In: Double, J.A., Thompson, M.J. (eds) Telomeres and Telomerase. Methods in Molecular Biology™, vol 191. Humana Press. https://doi.org/10.1385/1-59259-189-2:125
Download citation
DOI: https://doi.org/10.1385/1-59259-189-2:125
Publisher Name: Humana Press
Print ISBN: 978-0-89603-657-4
Online ISBN: 978-1-59259-189-3
eBook Packages: Springer Protocols