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地中海实蝇α-L-岩藻糖苷酶基因的克隆、序列鉴定及表达谱分析。

Cloning, sequence identification and expression profile analysis of α-L-fucosidase gene from the Mediterranean fruit fly Ceratitis capitata.

机构信息

Department of Biomolecular Sciences and Biotechnology, University of Milano, via Celoria 26, 20133 Milano, Italy.

出版信息

J Insect Physiol. 2011 Apr;57(4):452-61. doi: 10.1016/j.jinsphys.2011.01.007. Epub 2011 Jan 25.

Abstract

The Mediterranean fruit fly Ceratitis capitata (Diptera: Tephritidae) is one of the most destructive agricultural pests, a polyphagus insect of relevant economic importance and is widespread in many regions around the world. It is the best-studied fruit fly pest at genetic and molecular level and much has been learned on its ecology and behaviour. An α-L-fucosidase has been recently hypothesized to be involved in sperm-egg interactions in Drosophila melanogaster and in other Drosophila species. Here, a complete cDNA encoding a putative α-L-fucosidase of the medfly was amplified using the reverse polymerase chain reaction (RT-PCR) with degenerate based on the conserved coding sequence information of several insect α-L-fucosidases, cloned and sequenced (GenBank accession no. FJ177429). The coding region consisted of 1482 bp which encoded a 485-residues protein (named CcFUCA) with a predicted molecular mass of 56.1 kDa. The deduced protein sequence showed 75% amino acid identity to D. melanogaster α-L-fucosidase, and in fact the phylogenetic tree analysis revealed that CcFUCA had closer relationships with the α-L-fucosidases of drosophilid species. The tissue expression analysis indicated that CcFuca was expressed in a single transcript in all tissues, suggesting a ubiquitous localization pattern of the encoded protein. Our findings provide novel insights on a gene encoding a protein potentially involved in primary gamete interactions in C. capitata.

摘要

地中海实蝇 Ceratitis capitata(双翅目:实蝇科)是一种极具破坏性的农业害虫,是一种具有重要经济意义的杂食性昆虫,广泛分布于世界各地的许多地区。它是遗传和分子水平上研究得最好的实蝇害虫,在其生态学和行为学方面已经有了很多了解。最近有研究假设α-L-岩藻糖苷酶在黑腹果蝇和其他果蝇物种的精子-卵子相互作用中发挥作用。在这里,使用基于几种昆虫α-L-岩藻糖苷酶保守编码序列信息的简并引物,通过反转录聚合酶链反应(RT-PCR)扩增了 Medfly 中一个假定的α-L-岩藻糖苷酶的完整 cDNA,进行克隆和测序(GenBank 登录号 FJ177429)。编码区由 1482bp 组成,编码一个 485 个残基的蛋白质(命名为 CcFUCA),预测分子量为 56.1kDa。推导的蛋白质序列与黑腹果蝇的α-L-岩藻糖苷酶具有 75%的氨基酸同一性,实际上,系统发育树分析表明 CcFUCA 与果蝇属物种的α-L-岩藻糖苷酶具有更密切的关系。组织表达分析表明,CcFuca 在所有组织中都以单一转录本表达,表明编码蛋白具有普遍的定位模式。我们的研究结果为参与地中海实蝇初级配子相互作用的基因编码蛋白提供了新的见解。

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