Establishing an efficient Ac/Ds tagging system in rice:: large-scale analysis of Ds flanking sequences
Establishing an efficient Ac/Ds tagging system in rice:: large-scale analysis of Ds flanking sequences
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DOI：
10.1046/j.1365-313x.2003.01948.x
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发表时间：
2004-01-01
期刊：
影响因子：
7.2
通讯作者：
Ramachandran, S
中科院分区：
文献类型：
--
作者：
Kolesnik, T;Szeverenyi, I;Ramachandran, S
在水稻中成功建立了双元素激活/解离（Ac/Ds）基因诱捕体系。粳稻的履历。Nipponbare)产生一系列稳定的、非连锁的、单拷贝的Ds转座体。通过对4413个家庭的分析，估计Ds的生发转位频率平均为51%。对兄弟姐妹Ds转位模式的研究表明，79%的水稻至少有两个不同的插入，表明在水稻发育过程中发生了晚转位。对2057个Ds侧翼序列的分析表明，88%的Ds侧翼序列是唯一的，其余的在T-DNA内。这些插入在整个基因组中随机分布；然而，对4号和7号染色体有偏向，它们的插入数是预期的两倍。在7号染色体40kbp区域内发现了一个Ds插入热点。三分之一的Ds侧翼序列与蛋白质或水稻表达序列标签（ESTs）同源，证实了Ds优先转置到编码区。对1号染色体上200个Ds系的分析表明，72%的插入位于基因区。基于酵母人工染色体（YAC）的EST图谱锚定了800多个插入片段，结果表明，Ds优先转座到表达序列丰富的区域。高的萌发转座频率和兄弟间的独立转座表明该系统适合水稻大规模转座诱变。
A two-element Activator/Dissociation (Ac/Ds) gene trap system was successfully established in rice (Oryza sativa ssp. japonica cv. Nipponbare) to generate a collection of stable, unlinked and single-copy Ds transposants. The germinal transposition frequency of Ds was estimated as an average of 51% by analyzing 4413 families. Study of Ds transposition pattern in siblings revealed that 79% had at least two different insertions, suggesting late transposition during rice development. Analysis of 2057 Ds flanking sequences showed that 88% of them were unique, whereas the rest within T-DNA. The insertions were distributed randomly throughout the genome; however, there was a bias toward chromosomes 4 and 7, which had two times as many insertions as that expected. A hot spot for Ds insertions was identified on chromosome 7 within a 40-kbp region. One-third of Ds flanking sequences was homologous to either proteins or rice expressed sequence tags (ESTs), confirming a preference for Ds transposition into coding regions. Analysis of 200 Ds lines on chromosome 1 revealed that 72% insertions were found in genic region. Anchoring of more than 800 insertions to yeast artificial chromosome (YAC)-based EST map showed that Ds transposes preferentially into regions rich in expressed sequences. High germinal transposition frequency and independent transpositions among siblings show that the efficiency of this system is suitable for large-scale transposon mutagenesis in rice.