PMID: 9662067Jul 14, 1998Paper

Digital cloning: identification of human cDNAs homologous to novel kinases through expressed sequence tag database searching

Journal of Biomedical Science
H C ChenD Robinson

Abstract

Identification of novel kinases based on their sequence conservation within kinase catalytic domain has relied so far on two major approaches, low-stringency hybridization of cDNA libraries, and PCR method using degenerate primers. Both of these approaches at times are technically difficult and time-consuming. We have developed a procedure that can significantly reduce the time and effort involved in searching for novel kinases and increase the sensitivity of the analysis. This procedure exploits the computer analysis of a vast resource of human cDNA sequences represented in the expressed sequence tag (EST) database. Seventeen novel human cDNA clones showing significant homology to serine/threonine kinases, including STE-20, CDK- and YAK-related family kinases, were identified by searching EST database. Further sequence analysis of these novel kinases obtained either directly from EST clones or from PCR-RACE products confirmed their identity as protein kinases. Given the rapid accumulation of the EST database and the advent of powerful computer analysis software, this approach provides a fast, sensitive, and economical way to identify novel kinases as well as other genes from EST database.

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Citations

Jul 14, 1998·Journal of Biomedical Science·H J KungD Robinson
Jun 15, 1999·Parasitology Today·A C Ivens, J M Blackwell
Dec 28, 1999·Proceedings of the National Academy of Sciences of the United States of America·F ChenC P Austin
May 17, 2008·Nucleic Acids Research·Daniel AguilarFabien Campagne

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