Molecular cloning and characterization of complementary DNA encoding for ferredoxin-dependent glutamate synthase in maize leaf.
The sequence of ferredoxin-dependent glutamate synthase (EC 1.4.7.1) mRNA from maize has been determined. Complementary DNAs were isolated from a cDNA library of light-induced leaf poly(A)+ RNA constructed in an expression vector. An open reading frame beginning at an ATG codon at nucleotide 328 of the longest cDNA (5617-bases long) encoded 1616 amino acid residues. The amino terminus of the purified mature enzyme coincided with the cysteine residue at position 98 of the predicted sequence. This enzyme is homologous with the large subunit of Escherichia coli NADPH-dependent glutamate synthase having about 42% identical residues between the two proteins. The enzyme also contains a short region similar to a potential FMN-binding region of yeast flavocytochrome b2. The cDNA hybridizes to an RNA band about 5.5 kilobases whose steady-state level is markedly increased upon illumination of etiolated maize seedlings. Analysis of genomic DNA indicates the presence of a single-copy gene for ferredoxin glutamate synthase in maize.
Sakakibara
H
H
Department of Agricultural Chemistry, School of Agriculture, Nagoya University, Japan.
Watanabe
M
M
Hase
T
T
Sugiyama
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005288:Ferredoxins / Q000378:metabolism*; D005816:Genetic Complementation Test; D005970:Glutamate Synthase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D003313:Zea mays / Q000201:enzymology / Q000235:genetics*
D005288:Ferredoxins; D004247:DNA; D005970:Glutamate Synthase