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

  • Original Paper
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Journal of Biomedical Science

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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Authors and Affiliations

  1. Molecular and Genomic Medicine Division, National Health Research Institutes, 128 Yen-Chiu-Yuan Road, Sec. 2, 115, Taipei, Taiwan (ROC)

    Hua-Chien Chen & Hsing-Jien Kung

  2. Department of Molecular Biology and Microbiology, School of Medicine, Case Western Reserve University, Cleveland, Ohio, USA

    Hsing-Jien Kung & Dan Robinson

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  1. Hua-Chien Chen
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  2. Hsing-Jien Kung
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Chen, HC., Kung, HJ. & Robinson, D. Digital cloning: Identification of human cDNAs homologous to novel kinases through expressed sequence tag database searching. J Biomed Sci 5, 86–92 (1998). https://doi.org/10.1007/BF02258361

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