Cloning and Characterization of S-Adenosyl-L-methionine synthetase gene from Saccharomyces cerevisiae

  • Ko, Kwon-Hye (Department of Molecular Science and Technology, Ajou University) ;
  • Yoon, Gee-Sun (Department of Molecular Science and Technology, Ajou University) ;
  • Choi, Gi-Sub (Department of Molecular Science and Technology, Ajou University) ;
  • Suh, Joo-Won (Department of Biological Science, Institute of Bioscience and Biotechnology, Myongji University) ;
  • Ryu, Yeon-Woo (Department of Molecular Science and Technology, Ajou University)
  • Published : 2005.10.27

Abstract

S-Adenosyl-L-Methionine(SAM) has an important role for DNA methylation and cell signaling. SAM was synthesized from methionine and ATP by SAM synthetase and play an pivotal function in the primary and secondary metabolism of cells. Recent studies have revealed in the effect of SAM in case of morphological differentiation in both eukaryotes and prokaryotes. We isolated SAM gene from Saccharomyces cerevisiae and cloned it into expression vector for E. coli respectively. An 1.15 kb SAM-s gene fragment was isolated by Low-strigency PCR using ORF primer. By the analysed primary sequence deduced from DNA sequence, this gene included conserved domains similar with other well-known SAM synthetase. First of all, SAM synthetase gene cloned pGEM-T vector and subcloned into histidine tagging system to purify the expressed protein using metal chelating resin. Typical characteristic analysis of this enzyme is underway.

Keywords