Stolc Viktor, Gauhar Zareen, Mason Christopher, Halasz Gabor, van Batenburg Marinus F, Rifkin Scott A, Hua Sujun, Herreman Tine, Tongprasit Waraporn, Barbano Paolo Emilio, Bussemaker Harmen J, White Kevin P
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520, USA.
Science. 2004 Oct 22;306(5696):655-60. doi: 10.1126/science.1101312.
We used a maskless photolithography method to produce DNA oligonucleotide microarrays with unique probe sequences tiled throughout the genome of Drosophila melanogaster and across predicted splice junctions. RNA expression of protein coding and nonprotein coding sequences was determined for each major stage of the life cycle, including adult males and females. We detected transcriptional activity for 93% of annotated genes and RNA expression for 41% of the probes in intronic and intergenic sequences. Comparison to genome-wide RNA interference data and to gene annotations revealed distinguishable levels of expression for different classes of genes and higher levels of expression for genes with essential cellular functions. Differential splicing was observed in about 40% of predicted genes, and 5440 previously unknown splice forms were detected. Genes within conserved regions of synteny with D. pseudoobscura had highly correlated expression; these regions ranged in length from 10 to 900 kilobase pairs. The expressed intergenic and intronic sequences are more likely to be evolutionarily conserved than nonexpressed ones, and about 15% of them appear to be developmentally regulated. Our results provide a draft expression map for the entire nonrepetitive genome, which reveals a much more extensive and diverse set of expressed sequences than was previously predicted.
我们采用无掩膜光刻法制作了DNA寡核苷酸微阵列,其独特的探针序列覆盖了黑腹果蝇的整个基因组以及预测的剪接位点。我们测定了生命周期各主要阶段(包括成年雄性和雌性)蛋白质编码和非蛋白质编码序列的RNA表达情况。我们检测到93%的注释基因具有转录活性,并且内含子和基因间序列中41%的探针有RNA表达。与全基因组RNA干扰数据和基因注释的比较揭示了不同类别基因的可区分表达水平以及具有基本细胞功能的基因的较高表达水平。在约40%的预测基因中观察到了可变剪接,并且检测到5440种以前未知的剪接形式。与拟暗果蝇存在保守同线区域的基因具有高度相关的表达;这些区域的长度从10到900千碱基对不等。与未表达的序列相比,已表达的基因间和内含子序列在进化上更可能是保守的,并且其中约15%似乎受到发育调控。我们的结果提供了整个非重复基因组的表达图谱草案,该图谱揭示了比以前预测的更为广泛和多样的表达序列集。