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通过寡核苷酸微阵列和生物信息学分析探索犬钩虫和捻转血矛线虫之间的转录保守性。

Exploring transcriptional conservation between Ancylostoma caninum and Haemonchus contortus by oligonucleotide microarray and bioinformatic analyses.

作者信息

Cantacessi C, Loukas A, Campbell B E, Mulvenna J, Ong E K, Zhong W, Sternberg P W, Otranto D, Gasser R B

机构信息

Department of Veterinary Science, The University of Melbourne, 250 Princes Highway, Werribee, Victoria 3030, Australia.

出版信息

Mol Cell Probes. 2009 Feb;23(1):1-9. doi: 10.1016/j.mcp.2008.09.004. Epub 2008 Oct 11.

Abstract

In this study, we identified, using an established oligonucleotide microarray platform for the parasitic nematode Haemonchus contortus, transcripts that are 'conserved' between serum-activated and non-activated L3s of Ancylostoma caninum (aL3 and L3, respectively) and H. contortus by cross-species hybridization (CSH) at high stringency and conducted extensive bioinformatic analyses of the cross-hybridizing expressed sequence tags (ESTs). The microarray analysis revealed significant differential hybridization between aL3 and L3 for 32 molecules from A. caninum, of which 29 were shown to have homologues/orthologues in the free-living nematode Caenorhabditis elegans and/or A. caninum and the other three molecules had no homologues in current gene databases. 'Non-wildtype' RNAi phenotypes were recorded for 13 of the C. elegans homologues. A subset of 16 C. elegans homologues/orthologues (i.e. genes abce-1, act-2, C08H9.2, C55F2.1, calu-1, col-181, cpr-6, elo-2, asp-1, K07E3.4, rpn-2, sel-9, T28C12.4, hsb-1, Y57G11C.15 and ZK593.1) were predicted to interact genetically with a total of 156 (range 1-88) other genes. Gene ontology (GO) analysis of the interacting genes revealed that the most common subcategories were signal transduction (7%), intracellular protein transport and glycolysis (6.2%) within 'biological process'; nuclear (25.7%) and intracellular (19.8%) within 'cellular component'; and ATP-binding (14.4%) and protein-binding (8.4%) within 'molecular function'. The potential roles of key molecules in the two blood-feeding parasitic nematodes are discussed in relation to the known roles of their homologues/orthologues in C. elegans. The CSH approach used may provide a tool for the screening of genes conserved across a range of different taxa of parasites for which DNA microarray platforms are not available.

摘要

在本研究中,我们使用一种已建立的用于寄生线虫捻转血矛线虫的寡核苷酸微阵列平台,通过高严谨度的种间杂交(CSH),鉴定出犬钩口线虫血清激活的L3(aL3)和未激活的L3(L3)与捻转血矛线虫之间“保守”的转录本,并对杂交的表达序列标签(EST)进行了广泛的生物信息学分析。微阵列分析显示,犬钩口线虫的32个分子在aL3和L3之间存在显著的差异杂交,其中29个在自由生活线虫秀丽隐杆线虫和/或犬钩口线虫中有同源物/直系同源物,另外三个分子在当前基因数据库中没有同源物。对秀丽隐杆线虫的13个同源物记录了“非野生型”RNAi表型。预测16个秀丽隐杆线虫同源物/直系同源物(即基因abce-1、act-2、C08H9.2、C55F-II.1、calu-1、col-181、cpr-6、elo-2、asp-1、K07E3.4、rpn-2、sel-9、T28C12.4、hsb-1、Y57G11C.15和ZK593.1)总共与156个(范围1-88)其他基因发生遗传相互作用。对相互作用基因的基因本体论(GO)分析表明,最常见的子类别在“生物学过程”中为信号转导(7%)、细胞内蛋白质转运和糖酵解(6.2%);在“细胞成分”中为细胞核(25.7%)和细胞内(19.8%);在“分子功能”中为ATP结合(14.4%)和蛋白质结合(8.4%)。结合其在秀丽隐杆线虫中的同源物/直系同源物的已知作用,讨论了关键分子在这两种吸血寄生线虫中的潜在作用。所使用的CSH方法可能为筛选在一系列没有DNA微阵列平台的不同寄生虫类群中保守的基因提供一种工具。

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