Construction of gender-enriched cDNA archives for adult Oesophagostomum dentatum by suppressive-subtractive hybridization and a microarray analysis of expressed sequence tags
Construction of gender-enriched cDNA archives for adult Oesophagostomum dentatum by suppressive-subtractive hybridization and a microarray analysis of expressed sequence tags
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DOI：
10.1017/s0031182005009728
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发表时间：
2006-05-01
期刊：
影响因子：
2.4
通讯作者：
Gasser, R. B.
中科院分区：
文献类型：
--
作者：
Cottee, P. A.;Nisbet, A. J.;Gasser, R. B.
在本研究中，我们使用抑制性提取性杂交（SSH）构建了富含性别的cDNA文库，用于寄生线虫的成年阶段（SSH）的寄生线虫植物牙齿牙齿（SSH），来自雌性和男性的序列克隆（490）（490） ）并对EX序列标签（EST）进行了生物信息学和微阵列分析。总共获得了873台EST（440名男性和433名女性），获得了91％匕首微阵列分析的测序成功，代表了两个性别增强的库，揭示了其中391个的差异化杂交（75.8％）。其中，女性的220（56.3％）的信号强度明显高于男性，预计其中154个（70％）在秀丽隐杆线虫中具有同源性。这些同源物被预测参与关键的生物学过程，包括胚胎营养，配子发生，分子结合/转运或代谢，核酸合成和功能以及信号转导。在雄性O.牙本质中具有统计较高信号强度的171个EST中，有43.8％的EST在秀丽隐杆线虫中具有同源物。 The-E同源物包括主要的精子蛋白（MSP）或MSP样分子，角蛋白样分子，参与代谢，含PDZ结构蛋白的蛋白质，糖结合蛋白，蛋白质激酶，丝氨酸蛋白酶或蛋白酶抑制剂的分子，糖结合蛋白，蛋白酶蛋白质激酶，蛋白质蛋白质激酶，蛋白酶蛋白酶，蛋白质蛋白酶，蛋白质蛋白质蛋白质，蛋白质蛋白质蛋白质，蛋白酶蛋白质蛋白质，蛋白酶蛋白酶蛋白以及其他蛋白质，例如酶和各种推定的蛋白质。在秀丽隐杆线虫中没有已知同源物的287个EST（来自两个性别增强的cDNA库）中，有50个（17.4％）在其他线虫中具有同源物，其中8个具有同源性，在其他各种生物体中具有同源物，104个（36.2％）没有同源物（36.2％）对当前基因数据库的任何序列疾病。本研究为从富含性别的cDNA档案中分离和分子，生化和功能表征的分离，生化和功能表征奠定了基础。
In the present study, we constructed gender-enriched cDNA libraries for the adult stage of the parasitic nematode Oesophagostomum dentation (order Strongylida) using suppressive-subtractive hybridization (SSH), sequenced clones from the female-library and male-library (490 from each) and conducted bioinformatic and microarray analyses of the ex pressed sequence tags (ESTs). In total, 873 ESTs (440 male and 433 female) were obtained, achieving a sequencing Success Of 91%dagger Microarray analyses of 516 unique ESTs representing both gender-enriched libraries revealed differential hybridization for 391 of them (75.8%). Of these, 220 (56.3%) had significantly greater signal intensities in the female than in the male,,and 154 (70%) of these were predicted to have homologues in C. elegans. These homologues were predicted to be involved in key biological processes, including embryonic nutrition, gametogenesis, molecular binding/transport or metabolism, nucleic acid synthesis and function, and signal transduction. Of the 171 ESTs with statistically higher signal intensities in male O. dentatum, 43.8%, had homologues in C. elegans. The-e homologues included major sperm proteins (MSPs) or MSP-like molecules, keratin-like molecules, molecules involved in metabolism, PDZ domain-containing proteins, sugar binding proteins, protein kinases, serine proteases Or protease inhibitors, molecules involved in proteolysis and other proteins, Such as enzymes and various putative proteins. Of the 287 ESTs (from both gender-enriched cDNA libraries) with no known homologues in C. elegans, 50 (17.4%) had homologues in other nematodes, 8 had homologues in various other organisms and 104 (36.2%) had no homology to any sequence ill Current gene databases. The present study lays a foundation for the isolation and molecular, biochemical and functional characterization of selected genes from the gender-enriched cDNA archives established for O. dentatum.