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构建和分析雄性和雌性棉铃虫(Helicoverpa armigera)触角 cDNA 文库及潜在气味结合蛋白基因的表达分析。

Construction and analysis of cDNA libraries from the antennae of male and female cotton bollworms Helicoverpa armigera (Hübner) and expression analysis of putative odorant-binding protein genes.

机构信息

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.

出版信息

Biochem Biophys Res Commun. 2011 Apr 8;407(2):393-9. doi: 10.1016/j.bbrc.2011.03.032. Epub 2011 Mar 21.

Abstract

Two high-quality cDNA libraries were constructed from female and male antennae of the cotton bollworm Helicoverpa armigera (Hübner). The titers were approximately 2.0 × 10⁶ pfu/ml for females and 2.3 × 10⁶ pfu/ml for males, and this complies with the test requirement. From the libraries, 1750 male ESTs and 1640 female ESTs were sequenced and further analyzed. We identified 15 olfactory genes (12 are new), and 14 of them have the characteristic six conserved cysteine residues. With the exception of OBP9, all the genes were classified as classical OBP genes. By alignment and cluster analysis, the 14 classical OBPs were divided into pheromone binding protein (PBP) genes, odorant binding protein (OBP) genes, general odorant binding protein 1 (GOBP1) genes, general odorant binding protein 2 (GOBP2) genes and antennae binding protein (ABP) genes. Among these genes, we obtained three PBP genes (PBP1-PBP3) including two new PBP genes, one new ABP gene, nine new OBP genes (OBP1-OBP9), one known GOBP1 gene and one known GOBP2 gene. Furthermore, the expression patterns of these 14 classical OBP genes were investigated in various tissues by real-time quantitative polymerase chain reaction (qPCR). The results indicated that some OBP genes are expressed differently in different sexes and tissues, but most of them are highly expressed in antennae.

摘要

构建了两个高质量的棉铃虫(Helicoverpa armigera )雌雄触角 cDNA 文库。雌虫和雄虫的滴度分别约为 2.0×106 pfu/ml 和 2.3×106 pfu/ml,符合测试要求。从文库中测序并进一步分析了 1750 个雄性 EST 和 1640 个雌性 EST。鉴定出 15 个嗅觉基因(12 个为新基因),其中 14 个具有特征性的 6 个保守半胱氨酸残基。除 OBP9 外,所有基因均归类为经典 OBP 基因。通过比对和聚类分析,将 14 个经典 OBPs 分为信息素结合蛋白(PBP)基因、气味结合蛋白(OBP)基因、一般气味结合蛋白 1(GOBP1)基因、一般气味结合蛋白 2(GOBP2)基因和触角结合蛋白(ABP)基因。在这些基因中,我们获得了三个 PBP 基因(PBP1-PBP3),包括两个新的 PBP 基因、一个新的 ABP 基因、九个新的 OBP 基因(OBP1-OBP9)、一个已知的 GOBP1 基因和一个已知的 GOBP2 基因。此外,通过实时定量聚合酶链反应(qPCR)研究了这些 14 个经典 OBP 基因在不同组织中的表达模式。结果表明,一些 OBP 基因在不同性别和组织中的表达不同,但大多数在触角中高度表达。

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