Functional Genomics Center Zurich, ETH and University of Zurich, CH-8057 Zurich, Switzerland.
Plant Physiol. 2010 Feb;152(2):487-99. doi: 10.1104/pp.109.150185. Epub 2009 Dec 23.
Transcriptome profiling has become a routine tool in biology. For Arabidopsis (Arabidopsis thaliana), the Affymetrix ATH1 expression array is most commonly used, but it lacks about one-third of all annotated genes present in the reference strain. An alternative are tiling arrays, but previous designs have not allowed the simultaneous analysis of both strands on a single array. We introduce AGRONOMICS1, a new Affymetrix Arabidopsis microarray that contains the complete paths of both genome strands, with on average one 25mer probe per 35-bp genome sequence window. In addition, the new AGRONOMICS1 array contains all perfect match probes from the original ATH1 array, allowing for seamless integration of the very large existing ATH1 knowledge base. The AGRONOMICS1 array can be used for diverse functional genomics applications such as reliable expression profiling of more than 30,000 genes, detection of alternative splicing, and chromatin immunoprecipitation coupled to microarrays (ChIP-chip). Here, we describe the design of the array and compare its performance with that of the ATH1 array. We find results from both microarrays to be of similar quality, but AGRONOMICS1 arrays yield robust expression information for many more genes, as expected. Analysis of the ATH1 probes on AGRONOMICS1 arrays produces results that closely mirror those of ATH1 arrays. Finally, the AGRONOMICS1 array is shown to be useful for ChIP-chip experiments. We show that heterochromatic H3K9me2 is strongly confined to the gene body of target genes in euchromatic chromosome regions, suggesting that spreading of heterochromatin is limited outside of pericentromeric regions.
转录组谱分析已成为生物学中的常规工具。对于拟南芥(Arabidopsis thaliana),最常使用的是 Affymetrix ATH1 表达阵列,但它缺少参考菌株中存在的三分之一的所有注释基因。另一种选择是平铺阵列,但以前的设计不允许在单个阵列上同时分析两条链。我们引入了 AGRONOMICS1,这是一种新的 Affymetrix 拟南芥微阵列,它包含了两条基因组链的完整路径,平均每个 35 个碱基基因组序列窗口有一个 25 碱基探针。此外,新的 AGRONOMICS1 阵列包含了原始 ATH1 阵列中所有的完全匹配探针,允许无缝集成非常庞大的现有 ATH1 知识库。AGRONOMICS1 阵列可用于多种功能基因组学应用,例如可靠地表达分析超过 30000 个基因、检测选择性剪接以及与微阵列结合的染色质免疫沉淀(ChIP-chip)。在这里,我们描述了该阵列的设计,并将其性能与 ATH1 阵列进行了比较。我们发现两个微阵列的结果质量相似,但 AGRONOMICS1 阵列可获得更多基因的稳健表达信息,这是预期的。在 AGRONOMICS1 阵列上分析 ATH1 探针可产生与 ATH1 阵列非常相似的结果。最后,AGRONOMICS1 阵列被证明对 ChIP-chip 实验有用。我们表明,异染色质 H3K9me2 强烈局限于常染色质区域中靶基因的基因体,这表明异染色质的扩散在着丝粒区域之外是有限的。