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斜纹夜蛾保幼激素二醇激酶的克隆与结构特征分析

Cloning and structural characterization of juvenile hormone diol kinase in Spodoptera litura.

作者信息

Zeng Bao-Juan, Lv Yuan, Zhang Li-Li, Huang Li-Hua, Feng Qi-Li

机构信息

Guangzhou Key Laboratory of Insect Development Regulation and Application Research, School of Life Sciences, South China Normal University, Guangzhou, 510631, China.

出版信息

Insect Sci. 2016 Dec;23(6):819-828. doi: 10.1111/1744-7917.12236. Epub 2015 Jul 15.

Abstract

Juvenile hormone (JH) is one of the key insect hormones that regulate metamorphosis. Juvenile hormone diol kinase (JHDK) is an enzyme involved in JH metabolism and catalyzes JH diol to form a polar end product, JH diol phosphate that has no JH activity. In this study, a JHDK complementary DNA (cDNA) was cloned from Spodoptera litura and the structure and expression of the gene was characterized. The cDNA was 714 base pairs in length and encoded a protein of 183 amino acids with a molecular mass of 21 kDa and an isoelectric point of 4.55. Based on the structure, three putative calcium binding motifs and guanosine triphosphate-binding motifs were predicted in the protein. Modeling of the 3-D structure showed that the protein consisted of eight α-helixes linked with loops, with no β-sheets. The gene was expressed in the epidermis, fat body and midgut of fifth and sixth instar larvae. The expression level in the epidermis was lower than in the fat body and midgut. The gene was expressed at higher levels at the early stages than in the later stages of fifth and sixth instar midgut and fat body. The results suggest that this gene may be involved in the regulation of the JH titer in larvae of S. litura.

摘要

保幼激素(JH)是调节变态发育的关键昆虫激素之一。保幼激素二醇激酶(JHDK)是一种参与JH代谢的酶,催化JH二醇形成一种无JH活性的极性终产物——JH磷酸二醇。在本研究中,从斜纹夜蛾中克隆了JHDK互补DNA(cDNA),并对该基因的结构和表达进行了表征。该cDNA长度为714个碱基对,编码一个由183个氨基酸组成的蛋白质,分子量为21 kDa,等电点为4.55。基于该结构,预测该蛋白质中有三个假定的钙结合基序和鸟苷三磷酸结合基序。三维结构建模显示,该蛋白质由八个通过环连接的α螺旋组成,无β折叠。该基因在五龄和六龄幼虫的表皮、脂肪体和中肠中表达。在表皮中的表达水平低于脂肪体和中肠。在五龄和六龄中肠及脂肪体的早期阶段,该基因的表达水平高于后期阶段。结果表明,该基因可能参与斜纹夜蛾幼虫中JH滴度的调节。

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