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

Hua Chien Chen, Hsing-Jien Kung, Dan Robinson

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)86-92
Number of pages7
JournalJournal of Biomedical Science
Volume5
Issue number2
StatePublished - 1998
Externally publishedYes

Keywords

  • CDK
  • ERK
  • EST database
  • PKA
  • PKC
  • Protein kinase
  • STE-20
  • YAK

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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