Computational analysis and functional expression of ancestral copepod luciferase
Computational analysis and functional expression of ancestral copepod luciferase
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DOI：
10.1016/j.gene.2013.07.011
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发表时间：
2013-10-10
期刊：
影响因子：
3.5
通讯作者：
Shigeri, Yasushi
中科院分区：
文献类型：
--
作者：
Takenaka, Yasuhiro;Noda-Ogura, Akiko;Shigeri, Yasushi
最近，我们报道了11个桡足类的cDNA序列的总科Augaptiloidea在哲水蚤目。通过系统发育分析，将其分为两个类群：Metridinidae和Heterorhabdidae/Lucicutiidae。为了阐明进化过程，我们现在已经进一步从Augaptiloidea物种（Metridia asymmetrica，Metridia curticauda，Pleuromamma scutullata，Pleuromamma xiphias，Lucicutia ovaliformis和Heterorhabdus tanneri）中分离出12种桡足类酶。基于密码子的同义/非同义测试的正选择25个确定的桡足类的paclases表明，积极的达尔文选择在进化的异小杆科paclases操作，而两种类型的Metridinidae paclases多样化通过中性机制。通过对25个桡足类昆虫的编码氨基酸序列的计算机分析，我们推断出两个蛋白质序列是桡足类昆虫的祖先编码蛋白。它们在HEK 293细胞中表达，其中它们在细胞内裂解物和培养基中均表现出显著的荧光素酶活性，表明荧光素酶活性在这些桡足类物种进化多样化之前建立。C）2013 Elsevier B. V.保留所有权利。
We recently reported the cDNA sequences of 11 copepod luciferases from the superfamily Augaptiloidea in the order Calanoida. They were classified into two groups, Metridinidae and Heterorhabdidae/Lucicutiidae families, by phylogenetic analyses. To elucidate the evolutionary processes, we have now further isolated 12 copepod luciferases from Augaptiloidea species (Metridia asymmetrica, Metridia curticauda, Pleuromamma scutullata, Pleuromamma xiphias, Lucicutia ovaliformis and Heterorhabdus tanneri). Codon-based synonymous/nonsynonymous tests of positive selection for 25 identified copepod luciferases suggested that positive Darwinian selection operated in the evolution of Heterorhabdidae luciferases, whereas two types of Metridinidae luciferases had diversified via neutral mechanism. By in silico analysis of the decoded amino acid sequences of 25 copepod luciferases, we inferred two protein sequences as ancestral copepod luciferases. They were expressed in HEK293 cells where they exhibited notable luciferase activity both in intracellular lysates and cultured media, indicating that the luciferase activity was established before evolutionary diversification of these copepod species. C) 2013 Elsevier B.V. All rights reserved.