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从美洲棕榈象甲(Rhynchophorus palmarum L.)中鉴定和分子克隆假定的气味结合蛋白

Identification and molecular cloning of putative odorant-binding proteins from the American palm weevil, Rhynchophorus palmarum L.

作者信息

Nagnan-Le Meillour Patricia, François Marie-Christine, Jacquin-Joly Emmanuelle

机构信息

INRA, UR 258 Phytopharmacie et Médiateurs Chimiques, Route de Saint-Cyr, F-78026 Versailles Cedex, France.

出版信息

J Chem Ecol. 2004 Jun;30(6):1213-23. doi: 10.1023/b:joec.0000030273.77407.4d.

Abstract

We have identified and cloned the cDNAs encoding two odorant-binding proteins (OBPs) from the American palm weevil (APW) Rhynchophorus palmarum (Coleoptera, Curculionidae). Degenerate primers were designed from the N-terminal sequences and were used in polymerase chain reaction (PCR) in order to obtain full-length sequences in both males and females. In both sexes, two different cDNAs were obtained, encoding 123 and 115 amino acid-deduced sequences. Each sequence showed few amino acid differences between the sexes. The proteins were named RpalOBP2 and RpalOBP4 for male, RpalOBP2' and RpalOBP4' for female, with the types 2 and 4 presenting only 34% identities. These proteins shared high identity with previously described coleopteran OBPs. In native gels, RpalOBP2 clearly separated into two bands and RpalOBP4 into three bands, suggesting the presence of several conformational isomers. Thus, OBP diversity in this species may rely on both the presence of OBPs from different classes and the occurrence of isoforms for each OBP.

摘要

我们已经从美洲棕榈象甲(APW)红棕象甲(鞘翅目,象甲科)中鉴定并克隆了编码两种气味结合蛋白(OBP)的cDNA。根据N端序列设计简并引物,并用于聚合酶链反应(PCR),以获得雄性和雌性的全长序列。在两性中,获得了两种不同的cDNA,编码123和115个氨基酸推导序列。每个序列在两性之间显示出很少的氨基酸差异。这些蛋白质雄性命名为RpalOBP2和RpalOBP4,雌性命名为RpalOBP2'和RpalOBP4',2型和4型仅具有34%的同一性。这些蛋白质与先前描述的鞘翅目OBP具有高度同一性。在天然凝胶中,RpalOBP2明显分离为两条带,RpalOBP4分离为三条带,表明存在几种构象异构体。因此,该物种中的OBP多样性可能既依赖于不同类别的OBP的存在,也依赖于每种OBP的异构体的出现。

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