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一种用于曼氏血吸虫转录组分析的寡核苷酸微阵列及其在研究性别相关基因表达中的应用。

An oligonucleotide microarray for transcriptome analysis of Schistosoma mansoni and its application/use to investigate gender-associated gene expression.

作者信息

Fitzpatrick Jennifer M, Johnston David A, Williams Gary W, Williams Debbie J, Freeman Tom C, Dunne David W, Hoffmann Karl F

机构信息

Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QP, UK.

出版信息

Mol Biochem Parasitol. 2005 May;141(1):1-13. doi: 10.1016/j.molbiopara.2005.01.007.

DOI:10.1016/j.molbiopara.2005.01.007
PMID:15811522
Abstract

Global profiling transcriptomes of parasitic helminths offers the potential to simultaneously identify co-ordinately expressed genes, novel genetic programs and uniquely utilized metabolic pathways, which together provide an extensive and new resource for vaccine and drug discovery. We have exploited this post-genomic approach to fabricate the first oligonucleotide DNA microarray for gene expression analysis of the parasitic trematode Schistosoma mansoni. A total of 17,329 S. mansoni DNA sequences were used to design a microarray consisting of 7335 parasite elements or approximately 50% of this parasite's transcriptome. Here, we describe the design of this new microarray resource and its evaluation by extending studies into gender-associated gene expression in adult schistosomes. We demonstrate a high degree of reproducibility in detecting transcriptional differences among biologically replicated experiments and the ability of the microarray to distinguish between the expression of closely related gene family members. Importantly, for issues related to sexual dimorphism, labour division, gamete production and drug target discovery, 197 transcripts demonstrated a gender-biased pattern of gene expression in the adult schistosome, greatly extending the number of sex-associated genes. These data demonstrate the power of this new resource to facilitate a greater understanding into the biological complexities of schistosome development and maturation useful for identifying novel intervention strategies.

摘要

对寄生蠕虫进行全基因组转录组分析,有潜力同时鉴定出协同表达的基因、新的遗传程序以及独特利用的代谢途径,这些共同为疫苗和药物研发提供了丰富的新资源。我们利用这种后基因组方法构建了首个用于曼氏血吸虫基因表达分析的寡核苷酸DNA微阵列。总共17329条曼氏血吸虫DNA序列被用于设计一个由7335个寄生虫元件组成的微阵列,约占该寄生虫转录组的50%。在此,我们描述了这种新微阵列资源的设计,并通过对成年血吸虫性别相关基因表达的深入研究对其进行评估。我们证明了在检测生物重复实验之间的转录差异时具有高度的可重复性,以及微阵列区分密切相关基因家族成员表达的能力。重要的是,对于与性别二态性、分工、配子产生和药物靶点发现相关的问题,197个转录本在成年血吸虫中表现出性别偏向性的基因表达模式,极大地扩展了与性别相关的基因数量。这些数据证明了这种新资源有助于更深入了解血吸虫发育和成熟的生物学复杂性,从而有助于确定新的干预策略。

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