Molecular Cloning, Expression Pattern, and Phylogenetic Analysis of a Selenophosphate synthetase Gene From Antheraea pernyi (Lepidoptera: Saturniidae)
Molecular Cloning, Expression Pattern, and Phylogenetic Analysis of a Selenophosphate synthetase Gene From Antheraea pernyi (Lepidoptera: Saturniidae)
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DOI：
10.1603/an11068
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发表时间：
2011-11-01
影响因子：
2.3
通讯作者：
Liu, Yan-Qun
中科院分区：
文献类型：
--
作者：
Sun, Shou-Hui;Li, Yu-Ping;Liu, Yan-Qun
硒磷酸合成酶（Selenophosphate synthetase，Sps）是SelD基因的产物，能从ATP和硒化物中产生具有生物活性的硒供体化合物。本研究从重要经济昆虫柞蚕（Antheraea pernyi（Guerin-Meneville））（鳞翅目：天蚕科）中分离并鉴定了Sps基因。该cDNA序列全长1601 bp，开放阅读框为1209 bp，编码402个氨基酸，与果蝇（Drosophila melanogaster，Meigen）的同源性为87%。半定量RT-PCR分析表明，ApSps基因在家蚕4个发育阶段（卵、幼虫、蛹和成虫）和所有组织（血液、脂肪体、中肠、丝腺、体壁、精巢和卵巢）中都有转录，表明ApSps基因在家蚕的发育过程中起重要作用。pernyi。从数据库检索发现，Sps蛋白同源物存在于原核生物和真核生物中，包括细菌、真菌、无脊椎动物和脊椎动物，真核生物之间的氨基酸序列同源性为47-98%，表明它们在真核生物进化过程中高度保守。基于Sps蛋白同源序列的系统发育分析将已知的细菌、真菌、无脊椎动物和脊椎动物的Sps蛋白清晰地分开，与经典系统学的拓扑树一致，表明Sps蛋白序列在系统发育推断中具有潜在的价值。
Selenophosphate synthetase (Sps), the product of the SelD gene, produces a biologically active selenium donor compound from ATP and selenide. We have isolated and characterized the Sps gene from Antheraea pernyi (Guerin-Meneville) (Lepidoptera: Saturniidae), an economically important insect. The resulting 1601 bp cDNA sequence contains an open reading frame of 1209 bp encoding a polypeptide of 402 amino acids, with 87% sequence identity to that from Drosophila melanogaster (Meigen). Semiquantitative reverse transcription-polymerase chain reaction (PCR) analysis showed that the Sps gene was transcribed during four developmental stages (egg, larva, pupa, and adult) and in all the tissues tested (blood, fat body, midgut, silk glands, body wall, spermaries and ovaries), suggesting that ApSps plays an important role in the development of A. pernyi. From a database search, Sps protein homologs were found in prokaryotes and eukaryotes, including bacteria, fungi, invertebrates and vertebrates, with 47-98% amino acid sequence identities between eukaryotes, suggesting that they were highly conserved during the evolution of eukaryotes. Phylogenetic analysis, based on Sps protein homolog sequences, clearly separated the known bacterial, fungal, invertebrate and vertebrate Sps proteins, consistent with the topology tree of classical systematics, suggesting the potential value of the Sps protein sequence in phylogenetic inference.