Department of Biology & CESAM, University of Aveiro, Aveiro, Portugal.
PLoS One. 2012;7(4):e34266. doi: 10.1371/journal.pone.0034266. Epub 2012 Apr 27.
Enchytraeus albidus (Oligochaeta) is an ecologically relevant species used as standard test organisms for risk assessment. Effects of stressors in this species are commonly determined at the population level using reproduction and survival as endpoints. The assessment of transcriptomic responses can be very useful e.g. to understand underlying mechanisms of toxicity with gene expression fingerprinting. In the present paper the following is being addressed: 1) development of suppressive subtractive hybridization (SSH) libraries enriched for differentially expressed genes after metal and pesticide exposures; 2) sequencing and characterization of all generated cDNA inserts; 3) development of a publicly available genomic database on E. albidus. A total of 2100 Expressed Sequence Tags (ESTs) were isolated, sequenced and assembled into 1124 clusters (947 singletons and 177 contigs). From these sequences, 41% matched known proteins in GenBank (BLASTX, e-value ≤ 10(-5)) and 37% had at least one Gene Ontology (GO) term assigned. In total, 5.5% of the sequences were assigned to a metabolic pathway, based on KEGG. With this new sequencing information, an Agilent custom oligonucleotide microarray was designed, representing a potential tool for transcriptomic studies. EnchyBASE (http://bioinformatics.ua.pt/enchybase/) was developed as a web freely available database containing genomic information on E. albidus and will be further extended in the near future for other enchytraeid species. The database so far includes all ESTs generated for E. albidus from three cDNA libraries. This information can be downloaded and applied in functional genomics and transcription studies.
白正蚓(Oligochaeta)是一种生态相关物种,常被用作风险评估的标准试验生物。该物种的应激反应通常在种群水平上通过生殖和存活作为终点来确定。转录组反应的评估非常有用,例如通过基因表达指纹图谱来了解毒性的潜在机制。在本研究中,我们进行了以下研究:1)开发了针对金属和农药暴露后差异表达基因的抑制性消减杂交(SSH)文库;2)对所有生成的 cDNA 插入物进行测序和特征描述;3)开发了一个可公开获取的白正蚓基因组数据库。共分离出 2100 个表达序列标签(ESTs),进行测序并组装成 1124 个簇(947 个单克隆和 177 个基因簇)。从这些序列中,有 41%的序列与 GenBank 中的已知蛋白相匹配(BLASTX,e 值≤10(-5)),37%的序列至少有一个基因本体论(GO)术语。根据 KEGG,共有 5.5%的序列被分配到一个代谢途径。利用这些新的测序信息,设计了一个 Agilent 定制的寡核苷酸微阵列,代表了转录组研究的潜在工具。EnchyBASE(http://bioinformatics.ua.pt/enchybase/)是一个基于网络的免费数据库,包含了白正蚓的基因组信息,并将在不久的将来进一步扩展到其他蚓科物种。该数据库目前包含了从三个 cDNA 文库中为白正蚓生成的所有 ESTs。这些信息可以下载并应用于功能基因组学和转录研究。
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