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通过表达序列标签分析比较共生和非共生美丽艾氏海葵中的基因表达。

Comparative gene expression in the symbiotic and aposymbiotic Aiptasia pulchella by expressed sequence tag analysis.

作者信息

Kuo Jimmy, Chen Ming-Chyuan, Lin Chorng-Horng, Fang Lee-Shing

机构信息

Department of Planning and Research, National Museum of Marine Biology and Aquarium, Pingtung 944, Taiwan, ROC.

出版信息

Biochem Biophys Res Commun. 2004 May 21;318(1):176-86. doi: 10.1016/j.bbrc.2004.03.191.

Abstract

Intracellular symbiotic relationships are prevalent between cnidarians, such as corals and sea anemones, and the photosynthetic dinoflagellate symbionts. However, there is little understanding about how the genes express when the symbiotic relationship is set up. To characterize genes involved in this association, the endosymbiosis between sea anemone, Aiptasia pulchella, and dinoflagellate zooxanthellae, Symbiodinium spp., was employed as a model. Two complementary DNA (cDNA) libraries were constructed from RNA isolated from symbiotic and aposymbiotic A. pulchella. Using single-pass sequencing of cDNA clones, a total of 870 expressed sequence tags (ESTs) clones were generated from the two libraries: 474 from symbiotic animal and 396 from aposymbiotic animal. The initial ESTs consisted of 143 clusters and 231 singletons. A BLASTX search revealed that 147 unique genes had similarities with protein sequences available from databases; 120 of these clones were categorized according to their putative function. However, many ESTs could not assign functionally. The putative roles of some of the identified genes relative to endosymbiosis were discussed. This is the first report of the use of EST analysis to examine the gene expression in symbiotic and aposymbiotic states of the cnidarians. The systematic analysis of EST from this study provides a useful database for future investigations of the molecular mechanisms involved in algal-cnidarian symbiosis.

摘要

细胞内共生关系在刺胞动物(如珊瑚和海葵)与光合甲藻共生体之间普遍存在。然而,对于共生关系建立时基因如何表达却知之甚少。为了鉴定参与这种共生关系的基因,以海葵美丽艾氏海葵(Aiptasia pulchella)与甲藻虫黄藻(Symbiodinium spp.)之间的内共生关系为模型展开研究。从共生和非共生的美丽艾氏海葵中分离RNA,构建了两个互补DNA(cDNA)文库。通过对cDNA克隆进行单通道测序,从这两个文库中共获得了870个表达序列标签(EST)克隆:474个来自共生动物,396个来自非共生动物。最初的EST由143个聚类和231个单拷贝序列组成。BLASTX搜索显示,有147个独特基因与数据库中可获得的蛋白质序列具有相似性;其中120个克隆根据其推定功能进行了分类。然而,许多EST无法进行功能分配。讨论了一些已鉴定基因相对于内共生的推定作用。这是首次利用EST分析来检测刺胞动物共生和非共生状态下基因表达的报告。本研究对EST的系统分析为未来研究藻类 - 刺胞动物共生所涉及的分子机制提供了一个有用的数据库。

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