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利用二维凝胶电泳和质谱法检测臭虫(温带臭虫)精液中的蛋白质。

Detection of seminal fluid proteins in the bed bug, Cimex lectularius, using two-dimensional gel electrophoresis and mass spectrometry.

作者信息

Reinhardt K, Wong C H, Georgiou A S

机构信息

University of Sheffield, Department of Animal and Plant Sciences, Western Bank, Sheffield S10 2TN, UK.

出版信息

Parasitology. 2009 Mar;136(3):283-92. doi: 10.1017/S0031182008005362. Epub 2008 Dec 18.

Abstract

The global increase of the human parasite, the common bed bug Cimex lectularius, calls for specific pest control target sites. The bed bug is also a model species for sexual conflict theory which suggests that seminal fluids may be highly diverse. The species has a highly unusual sperm biology and seminal proteins may have unique functions. One-dimensional PAGE gels showed 40-50% band sharing between C. lectularius and another cimicid species, Afrocimex constrictus. However, adult, sexually rested C. lectularius males were found to store 5-7 microg of seminal protein and with only 60 microg of protein we obtained informative 2-D PAGE gels. These showed 79% shared protein spots between 2 laboratory populations, and more than half of the shared protein spots were detected in the mated female. Further analysis using liquid chromatography electrospray ionization tandem mass spectrometry revealed that 26.5% of the proteins had matches among arthropods in databases and 14.5% matched Drosophila proteins. These included ubiquitous proteins but also those more closely associated with reproduction such as moj 29, ubiquitin, the stress-related elongation factor EF-1 alpha, a protein disulfide isomerase and an antioxidant, Peroxiredoxin 6.

摘要

人类寄生虫温带臭虫(Cimex lectularius)在全球范围内数量增加,这就需要特定的害虫防治靶点。臭虫也是性冲突理论的模式物种,该理论认为精液可能具有高度多样性。该物种具有非常特殊的精子生物学特性,精液蛋白可能具有独特功能。一维聚丙烯酰胺凝胶电泳显示,温带臭虫与另一种臭虫物种——窄腹非洲臭虫(Afrocimex constrictus)之间有40%-50%的条带共享。然而,发现成年、性成熟的温带臭虫雄性储存有5-7微克的精液蛋白,我们仅用60微克的蛋白就获得了信息丰富的二维聚丙烯酰胺凝胶电泳图谱。这些图谱显示,两个实验室种群之间有79%的蛋白点是共享的,并且在交配后的雌性臭虫中检测到超过一半的共享蛋白点。使用液相色谱电喷雾电离串联质谱进行的进一步分析表明,26.5%的蛋白在数据库中的节肢动物中有匹配项,14.5%的蛋白与果蝇蛋白匹配。这些蛋白包括普遍存在的蛋白,也包括那些与繁殖关系更密切的蛋白,如moj 29、泛素、与应激相关的延伸因子EF-1α、一种蛋白二硫键异构酶和一种抗氧化剂——过氧化物酶6。

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