Insertional mutagenesis is a powerful tool for generating knockout mutations that facilitate associating biological functions with as yet uncharacterized open reading frames (ORFs) identified by genomic sequencing or represented in EST databases. We have generated a collection of Dissociation (Ds) transposon lines with insertions on all 5 Arabidopsis chromosomes. Here we report the insertion sites in 260 independent single-transposon lines, derived from four different Ds donor sites. We amplified and determined the genomic sequence flanking each transposon, then mapped its insertion site by identity of the flanking sequences to the corresponding sequence in the Arabidopsis genome database. This constitutes the largest collection of sequence-mapped Ds insertion sites unbiased by selection against the donor site. Insertion site clusters have been identified around three of the four donor sites on chromosomes 1 and 5, as well as near the nucleolus organizers on chromosomes 2 and 4. The distribution of insertions between ORFs and intergenic sequences is roughly proportional to the ratio of genic to intergenic sequence. Within ORFs, insertions cluster near the translational start codon, although we have not detected insertion site selectivity at the nucleotide sequence level. A searchable database of insertion site sequences for the 260 transposon insertion sites is available at http://sgio2.biotec.psu.edu/sr. This and other collections of Arabidopsis lines with sequence-identified transposon insertion sites are a valuable genetic resource for functional genomics studies because the transposon location is precisely known, the transposon can be remobilized to generate revertants, and the Ds insertion can be used to initiate further local mutagenesis.

中文翻译：

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拟南芥中序列和映射的Ds转座子插入位点的集合。

插入诱变是产生敲除突变的有力工具，该敲除突变促进将生物学功能与尚未被基因组测序鉴定或在EST数据库中表征的未表征的开放阅读框（ORF）相关联。我们已经生成了在所有5个拟南芥染色体上都有插入的解离（Ds）转座子系的集合。在这里，我们报告了从四个不同的Ds供体位点衍生的260个独立的单转座子系中的插入位点。我们扩增并确定了每个转座子侧翼的基因组序列，然后通过侧翼序列的身份将其插入位点映射到拟南芥基因组数据库中的相应序列。这构成了序列映射的Ds插入位点的最大集合，其通过针对供体位点的选择而没有偏倚。在染色体1和5的四个供体位点中的三个附近，以及在染色体2和4的核仁组织者附近，已经确定了插入位点簇。ORF和基因间序列之间的插入分布大致与基因对基因间序列。在ORF中，尽管我们尚未在核苷酸序列水平上检测到插入位点选择性，但插入簇聚集在翻译起始密码子附近。可在http://sgio2.biotec.psu.edu/sr上找到260个转座子插入位点的插入位点序列的可搜索数据库。带有序列鉴定的转座子插入位点的拟南芥品系和其他集合是功能基因组学研究的宝贵遗传资源，因为转座子的位置是众所周知的，