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 language | English |
|---|---|
| Pages (from-to) | 86-92 |
| Number of pages | 7 |
| Journal | Journal of Biomedical Science |
| Volume | 5 |
| Issue number | 2 |
| Publication status | Published - Jan 1 1998 |
| Externally published | Yes |
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Keywords
- CDK
- ERK
- EST database
- PKA
- PKC
- Protein kinase
- STE-20
- YAK
ASJC Scopus subject areas
- Endocrinology, Diabetes and Metabolism
- Molecular Biology
- Clinical Biochemistry
- Cell Biology
- Biochemistry, medical
- Pharmacology (medical)
Cite this
Digital cloning : Identification of human cDNAs homologous to novel kinases through expressed sequence tag database searching. / Chen, Hua Chien; Kung, Hsing Jien; Robinson, Dan.
In: Journal of Biomedical Science, Vol. 5, No. 2, 01.01.1998, p. 86-92.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Digital cloning
T2 - Identification of human cDNAs homologous to novel kinases through expressed sequence tag database searching
AU - Chen, Hua Chien
AU - Kung, Hsing Jien
AU - Robinson, Dan
PY - 1998/1/1
Y1 - 1998/1/1
N2 - 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.
AB - 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.
KW - CDK
KW - ERK
KW - EST database
KW - PKA
KW - PKC
KW - Protein kinase
KW - STE-20
KW - YAK
UR - http://www.scopus.com/inward/record.url?scp=0031747935&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0031747935&partnerID=8YFLogxK
M3 - Article
VL - 5
SP - 86
EP - 92
JO - Journal of Biomedical Science
JF - Journal of Biomedical Science
SN - 1021-7770
IS - 2
ER -