title: Expression, purification, and functional characterization of an N-terminal fragment of the tomato mosaic virus resistance protein Tm-1.
abstract: Tm-1, the protein product of Tm-1, a semidominant resistance gene of tomato, inhibits tomato mosaic virus (ToMV) replication by binding to ToMV replication proteins. Previous studies suggested the importance of the Tm-1 N-terminal region for its inhibitory activity; however, it has not been determined if the N-terminal region is sufficient for inhibition. Furthermore, the three-dimensional structure of Tm-1 has not been determined. In this study, an N-terminal fragment of Tm-1 (residues 1-431) as a fusion protein containing an upstream maltose-binding protein was expressed in E. coli Rosetta (DE3) cells at 30°C and then purified. The solubility of the fusion protein was greater when the cells were cultured at 30°C than when cultured at lower or higher temperatures. The purified N-terminal Tm-1 fragment from which the maltose-binding protein tag had been removed has inhibitory activity against ToMV RNA replication.
authors: Biomolecular Research Unit, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan.
authors: Masahiko
authors: M
authors: Kato
authors: Kazuhiro
authors: K
authors: Ishibashi
authors: Chihoko
authors: C
authors: Kobayashi
authors: Masayuki
authors: M
authors: Ishikawa
authors: Etsuko
authors: E
authors: Katoh
mesh_terms: D060467:Disease Resistance / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D018551:Solanum lycopersicum / Q000737:chemistry / Q000235:genetics* / Q000821:virology; D058619:Maltose-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D009029:Mosaic Viruses / Q000235:genetics / Q000472:pathogenicity; D010935:Plant Diseases / Q000235:genetics / Q000821:virology; D010940:Plant Proteins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D058619:Maltose-Binding Proteins; D010940:Plant Proteins