Analysis of expressed sequence tags from the four main developmental stages of Trypanosoma congolense.
Analysis of expressed sequence tags from the four main developmental stages of Trypanosoma congolense.
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DOI：
10.1016/j.molbiopara.2009.06.004
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发表时间：
2009-11
影响因子：
1.5
通讯作者：
Donelson, John E.
中科院分区：
文献类型：
--
作者：
Helm, Jared R.;Hertz-Fowler, Christiane;Aslett, Martin;Berriman, Matthew;Sanders, Mandy;Quail, Michael A.;Soares, Marcelo B.;Bonaldo, Maria F.;Sakurai, Tatsuya;Inoue, Noboru;Donelson, John E.
关键词：
刚果锥虫是非洲最重要的家畜病原体之一。利用刚毛虫生活史的3个主要阶段，即原环期、上成虫期和后环期的培养衍生寄生虫，以及从感染小鼠身上分离的血流期寄生虫，构建了阶段特异性的cDNA文库，并对每个文库中的表达序列标签(EST或cDNA克隆)进行了测序。在亚环文库中检测到13个编码不同变异表面糖蛋白(VSG)的EST簇，在血流文库中发现26个VSG EST簇，其中6个被亚环文库共享。少见的VSG EST存在于表鞭毛体文库中，而在原环状文库中未检测到。编码催化氧化磷酸化和氨基酸代谢的酶的EST在原环期和表鞭毛体阶段的含量大约是杂环期和血流期的两倍。相反，编码糖酵解、柠檬酸循环和核苷酸代谢的EST在所有四个发育阶段都大致相同。半胱氨酸蛋白酶、激酶和磷酸酶是ESTs中含量最丰富的酶类。所有四个文库都包含布氏毛滴虫和克鲁兹毛滴虫基因组中没有的贡氏毛滴虫特异的表达序列。从后循环和血流阶段构建了归一化的cDNA文库，并发现进一步丰富了弓形虫特异的EST。鉴于养殖的贡氏锥虫比其他非洲锥虫物种具有实验上的优势，这些EST为进一步研究这四个发育阶段的分子特性提供了基础，特别是难以获得大量生物的上胚体和中环阶段。可从以下网址获得T.congolense EST数据库：http://www.sanger.ac.uk/Projects/T_congolense/EST_index.shtml.
Trypanosoma congolense is one of the most economically important pathogens of livestock in Africa. Culture-derived parasites of each of the three main insect stages of the T. congolense life cycle, i.e., the procyclic, epimastigote and metacyclic stages, and bloodstream stage parasites isolated from infected mice, were used to construct stage-specific cDNA libraries and expressed sequence tags (ESTs or cDNA clones) in each library were sequenced. Thirteen EST clusters encoding different variant surface glycoproteins (VSGs) were detected in the metacyclic library and twenty-six VSG EST clusters were found in the bloodstream library, six of which are shared by the metacyclic library. Rare VSG ESTs are present in the epimastigote library, and none were detected in the procyclic library. ESTs encoding enzymes that catalyze oxidative phosphorylation and amino acid metabolism are about twice as abundant in the procyclic and epimastigote stages as in the metacyclic and bloodstream stages. In contrast, ESTs encoding enzymes involved in glycolysis, the citric acid cycle and nucleotide metabolism are about the same in all four developmental stages. Cysteine proteases, kinases and phosphatases are the most abundant enzyme groups represented by the ESTs. All four libraries contain T. congolense-specific expressed sequences not present in the T. brucei and T. cruzi genomes. Normalized cDNA libraries were constructed from the metacyclic and bloodstream stages, and found to be further enriched for T. congolense-specific ESTs. Given that cultured T. congolense offers an experimental advantage over other African trypanosome species, these ESTs provide a basis for further investigation of the molecular properties of these four developmental stages, especially the epimastigote and metacyclic stages for which it is difficult to obtain large quantities of organisms. The T. congolense EST databases are available at: http://www.sanger.ac.uk/Projects/T_congolense/EST_index.shtml.
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