title: Cloning, characterization and prokaryotic expression of cytosolic malate dehydrogenase from Oryza sativa.
abstract: cDNA sequences of malate dehydrogenase (MDH) were cloned from various species, and MDH was identified to play an important role in cell energy metabolism. Here, we present the isolation and characterization of its homologue (OscMDH) in Oryza sativa. Comparison of the results to the genome details indicated that OscMDH consisted of seven exons. Sequence alignment showed that the deduced amino acid sequence of OscMDH shared a significant similarity with cMDH protein in Zea mays, as well as with other cMDH gene products. The different expression patterns of OscMDH in different tissues revealed that OscMDH mRNA was highly transcribed in either young panicle or immature seed, while its abundance was much low in green leaf and root. A nearly 56-kDa fusion protein generated by adding a Trx-His-tag at the N-terminal of OscMDH was induced by IPTG in Escherichia coli BL21 and an obvious MDH activity was detected in the protein by native PAGE analysis. All these results suggest that OscMDH encodes a cytosolic MDH in rice.
authors.affiliation: Institute of Genetics, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shangai, People's Republic of China.
authors.forename: Chang-Fa
authors.initials: CF
authors.lastname: Lin
authors.forename: Rui-Hua
authors.initials: RH
authors.lastname: Jiang
authors.forename: Li-Zhi
authors.initials: LZ
authors.lastname: Jiang
authors.forename: Xiao-Yin
authors.initials: XY
authors.lastname: Qian
authors.forename: Kotb
authors.initials: K
authors.lastname: Attia
authors.forename: Jin-Shui
authors.initials: JS
authors.lastname: Yang
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D008291:Malate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009928:Organ Specificity; D012275:Oryza / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D012639:Seeds / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D000199:Actins; D008291:Malate Dehydrogenase