Cloning, characterization and prokaryotic expression of cytosolic malate dehydrogenase from Oryza sativa.
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.
Lin
Chang-Fa
CF
Institute of Genetics, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shangai, People's Republic of China.
Jiang
Rui-Hua
RH
Jiang
Li-Zhi
LZ
Qian
Xiao-Yin
XY
Attia
Kotb
K
Yang
Jin-Shui
JS
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
D000199:Actins; D008291:Malate Dehydrogenase