Molecular cloning, expression pattern and phylogenetic analysis of the will die slowly gene from the Chinese oak silkworm, Antheraea pernyi
Molecular cloning, expression pattern and phylogenetic analysis of the will die slowly gene from the Chinese oak silkworm, Antheraea pernyi
复制标题
柞蚕慢死基因的分子克隆、表达模式及系统发育分析
DOI：
10.1007/s11033-010-0495-2
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发表时间：
2011-08-01
影响因子：
2.8
通讯作者：
Wang, ZhenDong
中科院分区：
文献类型：
--
作者：
Li, Yuping;Wang, Huan;Wang, ZhenDong
将缓慢死亡（wds）基因编码的WD重复蛋白与七个重复已被确定在果蝇。本研究从中国柞蚕（Antheraea pernyi）中分离并鉴定了wds基因。该cDNA序列全长1733 bp，开放阅读框为1041 bp，编码346个氨基酸，与D.黑腹菌RT-PCR分析表明，wds基因在家蚕的4个发育阶段和所有组织中都有转录，这与基于EST资源和全基因组芯片信息在家蚕中观察到的结果一致。A.温度胁迫下，柞蚕wds基因的表达量与对照相比没有显著的下调或上调，表明该基因可能不参与温度胁迫耐受。通过数据库检索发现，wds蛋白在真菌、植物、无脊椎动物和脊椎动物等多种真核生物中存在同源性，氨基酸序列同源性在50-93%之间，表明其在真核生物进化过程中高度保守。基于wds蛋白同源序列的系统发育分析将已知的真菌、植物、无脊椎动物和脊椎动物清晰地分开，与经典系统学上的拓扑树一致，表明了wds蛋白在真核生物系统发育推断中的潜在价值。在脊椎动物中，通过序列比对和系统发育分析，也确定了两种明显的wds蛋白。
The will die slowly (wds) gene coding for a WD-repeat protein with seven repeats has been characterized in Drosophila melanogaster. In this paper, the wds gene was isolated and characterized from the Chinese oak silkworm, Antheraea pernyi (Lepidoptera: Saturniidae). The obtained 1733 bp cDNA sequence contains an open reading frame of 1041 bp encoding a polypeptide of 346 amino acids, with 85% sequence identity to that from D. melanogaster. RT-PCR analysis showed that the wds gene was transcribed during four developmental stages and in all the tissues tested, consistent with the result observed in Bombyx mori based on EST resources and genome-wide microarray information. The mRNA expression level of the A. pernyi wds gene was not significantly down- or up- regulated under temperature stress compared to the control, indicating that it may be not involved in temperature stress tolerance. In search of database, the wds protein homologues were found in various kinds of eukaryotes, including fungi, plants, invertebrates and vertebrates, with 50-93% amino acid sequence identities between them, suggesting that they are highly conserved during the evolution of eukaryotes. Phylogenetic analysis based on the wds protein homologue sequences clearly separated the known fungi, plants, invertebrates and vertebrates, consistent with the topology tree on the classical systematics, suggesting the potential value of wds protein in eukaryotic phylogenetic inference. In vertebrates, two apparent types of the wds proteins were also defined by sequence alignment and phylogenetic analysis.