Cloning and Characterization of Recombinant Substrate for Phophorylation of cdc2 Protein Kinase using a Radioisotope Detection Technique for the Development of a Microarray
Cell cycle-dependent protein kinases (CDKs) regulates the cells entry into mitosis during the transition from G2 to M phase, by phosphorylating a group of key proteins including RNA polymerase II and histone H1. A recombinant substrate for the enzyme was expressed as a recombinant protein from a plasmid pMFS-cdc2 in E. coli, and it was purified. In this study, we present a strategy for the detection of a phosphorylation of a substrate catalysed by cdc2 protein kinase by using a radioisotope detection technique in microarrays. We conclude that the use of Mep45-fused substrate in developing a protein biochip for the detection of the activity of cdc2 protein kinase by using a RI detection technique is highly economical and can be used for both high throughput screening and studying protein-protein interactions. Keywords: Cdc2 protein kinase, Gene cloning, Microarrays, Radioisotope detection
저자
Kyong-Cheol Ko [ Radiation Research Division for Biotechnology, Korea Atomic Energy Research Institute ]
Mi Hee Choi [ Radiation Research Division for Biotechnology, Korea Atomic Energy Research Institute ]
Sang Hyun Park [ Radiation Research Division for Biotechnology, Korea Atomic Energy Research Institute ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
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