Molecular cloning and characterization of complementary DNA encoding for ferredoxin-dependent glutamate synthase in maize leaf.
Molecular cloning and characterization of complementary DNA encoding for ferredoxin-dependent glutamate synthase in maize leaf.
复制标题
编码玉米叶中铁氧还蛋白依赖性谷氨酸合酶的互补 DNA 的分子克隆和表征。
DOI：
10.1016/s0021-9258(18)52204-3
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Tatsuo Sugiyama
中科院分区：
文献类型：
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作者：
Hitoshi Sakakibara;Mitsuru Watanabe;Toshiharu HaseS;Tatsuo Sugiyama
测定了玉米铁氧还蛋白依赖性谷氨酸合酶（EC 1.4.7.1）mRNA的序列。从构建在表达载体中的光诱导叶poly（A）+ RNA的cDNA文库中分离互补DNA。最长的cDNA（5617个碱基长）的第328位核苷酸的ATG密码子开始的开放阅读框架编码1616个氨基酸残基。纯化的成熟酶的氨基末端与预测序列的位置98处的半胱氨酸残基一致。该酶与大肠杆菌NADPH依赖性谷氨酸合酶的大亚基同源，两种蛋白质之间具有约42%的相同残基。该酶还含有一个类似于酵母flavocytochrome b2的潜在FMN结合区的短区域。cDNA杂交到约5.5个酶的RNA带，其稳态水平在黄化玉米幼苗光照后显著增加。基因组DNA分析表明，玉米中存在铁氧还蛋白谷氨酸合酶的单拷贝基因。
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.