S-Adenosylmethionine (SAM) is a cofactor for methyl group transfer in both prokaryotic and eukaryotic organisms and produced by S-Adenosylmethionine synthetase.1) To investigate the characterization of SAM-s from Pichia ciferrii for further applications, we cloned SAM-s from P. ciferrii by RACE, RNA capfishing and DNA walking. Cloned SAM-s encodesa 383 amino acids and 42KDa of size. After comparison of known other SAM-s sequences, cloned SAM-s showed about 90% of match with yeast SAM-s. The cloned SAM-s also contains consensus ATP-binding motif (G-X-G-X-X-G) and metal binding site (23D and 279D for Mg2+ and 51G for K+) which are located in other known SAM-s genes.2) Cloned SAM-s was inserted into E. coli expression vector to confirm their activity. After transformation, SDS-PAGE analysis was performed and we confirmed that cloned SAM-s was successfully expressed. However, no activity of cloned SAM- in E. coli was observed. Thus, cloned SAM-s was expressed in Pichia sp. for confirmation. The enzyme activity will be measured and result will be discussed.
저자
Sang-Young Yoon [ Sang-YoungYoon, | Dept of Molecular Science and Technology, Ajou University ]
Won-Kyu Lee [ Dept of Molecular Science and Technology, Ajou University ]
Yeon-Woo Ryu [ Dept of Molecular Science and Technology, Ajou University ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
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