Cloning of nicotianamine synthase genes from Arabidopsis thaliana
Cloning of nicotianamine synthase genes from Arabidopsis thaliana
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DOI：
10.1080/00380768.1999.10414350
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发表时间：
1999-12
影响因子：
2
通讯作者：
Kazuya Suzuki;K. Higuchi;H. Nakanishi;N. Nishizawa;S. Mori
中科院分区：
文献类型：
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作者：
Kazuya Suzuki;K. Higuchi;H. Nakanishi;N. Nishizawa;S. Mori
摘要烟酰胺合酶（NAS）催化S-腺苷甲硫氨酸三聚形成一分子烟酰胺（NA）。NA存在于所有植物中;它螯合金属阳离子，并被认为在植物中的金属稳态中起作用。此外，在禾本科单子叶植物中，NA是生物合成mugineic acid家族植物铁载体（MAs）的必需中间体。为了鉴定双子叶植物中编码NAS的基因，使用我们最近分离的大麦NAS基因（HvNAS）的核苷酸序列搜索拟南芥数据库。我们在数据库中发现了几个EST和三个基因组序列与HvNAS高度同源。根据这些核苷酸序列和HvNAS的核苷酸序列，我们设计了2组引物来分离拟南芥中的NAS直系同源物，并成功获得了3个编码AtNAS的DNA克隆（AtNAS 1、2和3）。这些克隆在大肠杆菌中得到表达，其蛋白产物显示出N-末端氨基酸残基。
Abstract Nicotianamine synthase (NAS) catalyzes the trimerization of S-adenosylmethionine to form one molecule of nicotianamine (NA). NA is present in all the plants; it chelates metal cations, and is considered to play a role in metal homeostasis in plants. Moreover, in graminaceous monocotyledonous plants, NA is an essential intermediate in the biosynthesis of mugineic acid family phytosiderophores (MAs). In order to identify the gene encoding NAS in dicotyledonous plants, Arabidopsis thaliana databases were searched using the nucleotide sequence of the NAS gene from barley (HvNAS), which we have recently isolated. We found several ESTs and three genomic sequences highly homologous to HvNAS in the databases. Based on these nucleotide sequences and that of HvNAS, we designed 2 sets of primers to isolate the NAS orthologues in Arabidopsis and succeeded in obtaining three DNA clones encoding AtNAS (AtNAS1, 2, and 3). These clones were expressed in Escherichia coli and their protein products displayed the N...