원문정보
초록
영어
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.