title: Digital cloning: identification of human cDNAs homologous to novel kinases through expressed sequence tag database searching.
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.
authors.affiliation: Molecular and Genomic Medicine Division, National Health Research Institutes, Taipei, Taiwan, ROC.
authors.forename: H C
authors.initials: HC
authors.lastname: Chen
authors.forename: H J
authors.initials: HJ
authors.lastname: Kung
authors.forename: D
authors.initials: D
authors.lastname: Robinson
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics*; D016208:Databases, Factual; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D011494:Protein Kinases / Q000235:genetics*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D011494:Protein Kinases