Building a human kinase gene repository: Bioinformatics, molecular cloning, and functional validation Academic Article uri icon

Overview

MeSH Major

  • Cloning, Molecular
  • Computational Biology
  • Databases, Genetic
  • Protein Kinases

abstract

  • Kinases catalyze the phosphorylation of proteins, lipids, sugars, nucleosides, and other important cellular metabolites and play key regulatory roles in all aspects of eukaryotic cell physiology. Here, we describe the mining of public databases to collect the sequence information of all identified human kinase genes and the cloning of the corresponding ORFs. We identified 663 genes, 511 encoding protein kinases, and 152 encoding nonprotein kinases. We describe the successful cloning and sequence verification of 270 of these genes. Subcloning of this gene set in mammalian expression vectors and their use in high-throughput cell-based screens allowed the validation of the clones at the level of expression and the identification of previously uncharacterized modulators of the survivin promoter. Moreover, expressions of the kinase genes in bacteria, followed by autophosphorylation assays, identified 21 protein kinases that showed autocatalytic activity. The work described here will facilitate the functional assaying of this important gene family in phenotypic screens and their use in biochemical and structural studies.

publication date

  • June 7, 2005

Research

keywords

  • Academic Article

Identity

Language

  • eng

PubMed Central ID

  • PMC1149441

Digital Object Identifier (DOI)

  • 10.1073/pnas.0503141102

PubMed ID

  • 15928075

Additional Document Info

start page

  • 8114

end page

  • 9

volume

  • 102

number

  • 23