Li Yong, Rosso Mario G, Ulker Bekir, Weisshaar Bernd
Max Planck Institute for Plant Breeding Research, Carl-von-Linne-Weg 10, D-50829 Cologne, Germany.
Genomics. 2006 May;87(5):645-52. doi: 10.1016/j.ygeno.2005.12.010. Epub 2006 Feb 20.
Large collections of sequence-indexed T-DNA insertion mutants are invaluable resources for plant functional genomics. Flanking sequence tag (FST) data from these collections indicated that T-DNA insertions are not randomly distributed in the Arabidopsis thaliana genome and that there are still a fairly high number of annotated genes without T-DNA insertions. We have analyzed FST data from the FLAGdb, GABI-Kat, and SIGnAL mutant populations. The lack of detectable transcriptional activity and the absence of suitable restriction sites were among the reasons genes are not covered by insertions. Additionally, a refined analysis of FSTs to genes with annotated noncoding regions showed that transcription initiation and polyadenylation site regions of genes are favored targets for T-DNA integration. These findings have implications for the use of T-DNA in saturation mutagenesis and for our chances to find a useful knockout allele for every gene.
大量经序列索引的T-DNA插入突变体文库是植物功能基因组学的宝贵资源。这些文库中的侧翼序列标签（FST）数据表明，T-DNA插入在拟南芥基因组中并非随机分布，而且仍有相当数量的注释基因没有T-DNA插入。我们分析了来自FLAGdb、GABI-Kat和SIGnAL突变体群体的FST数据。缺乏可检测到的转录活性以及没有合适的限制性酶切位点是基因未被插入覆盖的部分原因。此外，对具有注释非编码区的基因的FST进行的精细分析表明，基因的转录起始和聚腺苷酸化位点区域是T-DNA整合的偏好靶点。这些发现对于T-DNA在饱和诱变中的应用以及我们为每个基因找到有用的敲除等位基因的机会具有重要意义。
