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含有保守结构域WD40的家蚕BmPLA2影响家蚕脂肪体组织的代谢功能。

BmPLA2 containing conserved domain WD40 affects the metabolic functions of fat body tissue in silkworm, Bombyx mori.

作者信息

Orville Singh Chabungbam, Xin Hu-Hu, Chen Rui-Ting, Wang Mei-Xian, Liang Shuang, Lu Yan, Cai Zi-Zheng, Miao Yun-Gen

机构信息

Institute of Sericulture and Apiculture, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.

出版信息

Insect Sci. 2016 Feb;23(1):28-36. doi: 10.1111/1744-7917.12189. Epub 2015 Feb 3.

Abstract

PLA2 enzyme hydrolyzes arachidonic acid, and other polyunsaturated fatty acids, from the sn-2 position to release free arachidonic acid and a lysophospholipid. Previous studies reported that the PLA2 in invertebrate organisms participates in lipid signaling molecules like arachidonic acid release in immune-associated tissues like hemocytes and fat bodies. In the present study, we cloned the BmPLA2 gene from fat body tissue of silkworm Bombyx mori, which has a total sequence of 1.031 kb with a 31.90 kDa protein. In silico results of BmPLA2 indicated that the protein has a putative WD40 conserved domain and its phylogeny tree clustered with Danaus plexippus species. We investigated the transcriptional expression in development stages and tissues. The highest expression of BmPLA2 was screened in fat body among the studied tissues of third day fifth instar larva, with a high expression on third day fifth instar larva followed by a depression of expression in the wandering stage of the fifth instar larva. The expression of BmPLA2 in female pupa was higher than that of male pupa. Our RNAi-mediated gene silencing results showed highest reduction of BmPLA2 expression in post-24 h followed by post-48 and post-72 h. The BmPLA2-RNAi larvae and pupa could be characterized by pharate adult lethality and underdevelopment. The phenotypic characters of fat body cells in RNAi-induced larva implied that BmPLA2 affects the metabolic functions of fat body tissue in silkworm Bombyx mori.

摘要

磷脂酶A2(PLA2)可将花生四烯酸及其他多不饱和脂肪酸从sn-2位水解,释放出游离的花生四烯酸和溶血磷脂。先前的研究报道,无脊椎动物体内的PLA2参与脂质信号分子的产生,如在血细胞和脂肪体等免疫相关组织中释放花生四烯酸。在本研究中,我们从家蚕脂肪体组织中克隆了BmPLA2基因,其全长序列为1.031 kb,编码一个31.90 kDa的蛋白质。BmPLA2的电子克隆结果表明,该蛋白具有一个假定的WD40保守结构域,其系统发育树与黑脉金斑蝶聚类。我们研究了其在发育阶段和组织中的转录表达。在五龄第三天幼虫的各研究组织中,BmPLA2在脂肪体中的表达最高,在五龄第三天幼虫中表达较高,随后在五龄幼虫化蛹阶段表达下降。BmPLA2在雌蛹中的表达高于雄蛹。我们的RNA干扰介导的基因沉默结果显示,在24小时后BmPLA2表达的降低最为显著,其次是48小时和72小时后。BmPLA2-RNA干扰的幼虫和蛹表现为化蛹成虫致死和发育不全。RNA干扰诱导的幼虫脂肪体细胞的表型特征表明,BmPLA2影响家蚕脂肪体组织的代谢功能。

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