The expression pattern of four odorant-binding proteins in male and female silk moths, Bombyx mori
The expression pattern of four odorant-binding proteins in male and female silk moths, Bombyx mori
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DOI：
10.1007/s11068-005-5054-8
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发表时间：
2005-03-01
期刊：
影响因子：
--
通讯作者：
Steinbrecht, RA
中科院分区：
文献类型：
--
作者：
Maida, R;Mameli, M;Steinbrecht, RA
将家蚕4种气味结合蛋白--信息素结合蛋白（PBP）、一般气味结合蛋白1（GOBP 1）、一般气味结合蛋白2（GOBP 2）和触角结合蛋白X（ABPX）在大肠杆菌中表达。大肠杆菌并用于制备多克隆抗血清。触角匀浆的免疫印迹表明，这些抗血清是特异性的。免疫印迹分析和免疫细胞化学标记实验表明，重组PBP和GOBP 2的抗B. Mori分别给出了与针对天蚕的天然PBP和GOBP 2的血清相同的结果，从而证实了用后者获得的早期结果。用四种抗血清标记不同类型感器的连续横截面，发现雄性和雌性感器的标记模式不同。Alciddea和S. basiconica。很长。雄性和雌性的拟蚤属均为同一标记类型，而短拟蚤属则为同一标记类型。拉斯代亚山intermedia和S. basiconica观察到了各种各样的标记模式，有些比其他的更常见。很长。在雄性中均匀地调谐到信息素成分bombykol和bombykal，所有强烈表达PBP;对其他三种气味结合蛋白的抗血清标记几乎没有高于背景，只有在一些毛发中GOBP 1表达得更强烈。很长。女性，具体响应芳樟醇和苯甲酸，表现出不同的标签模式。在这里，我们观察到用抗GOBP 2抗体的强标记和用抗GOBP 1的中等标记，有时用抗ABPX。S.在两种性别的basiconica最常见的GOBP 1和GOBP 2共表达，但偶尔发现其他模式，其中一些显示PBP表达，也在女性。在短的标签类型的巨大变化。拉斯代亚山intermedia和S. basiconica提出了一个类似的多种功能亚型观察到的植物气味敏感感器的其他蛾类。
Four recombinant odorant-binding proteins of Bombyx mori, pheromone-binding protein (PBP), general odorant-binding protein 1 (GOBP1), general odorant-binding protein 2 (GOBP2) and antennal binding protein X (ABPX), were expressed in E. coli and used to raise polyclonal antisera. Immunoblots of antennal homogenates showed that these antisera were specific. In Western blot analysis and immunocytochemical labelling experiments, the sera against recombinant PBP and GOBP2 of B. mori gave identical results as sera against native PBP and GOBP2 of Antheraea polyphemus, respectively, thus confirming earlier results obtained with the latter. Labelling consecutive cross sections of various sensillum types with all four antisera revealed different labelling patterns in male and female sensilla (s.) trichodea and s. basiconica. Long s. trichodea in males and females represented uniform labelling types, whereas for short s. trichodea, s. intermedia, and s. basiconica a great variety of labelling patterns was observed, some being more common than others. Long s. trichodea, which in males are uniformly tuned to the pheromone components bombykol and bombykal, all strongly expressed PBP; labelling with antisera against the other three odorant-binding proteins hardly was above background, only in some hairs GOBP1 was expressed somewhat more strongly. Long s. trichodea of females, which respond specifically to linalool and benzoic acid, showed a different labelling pattern. Here, we observed strong labelling with antibodies against GOBP2 and medium labelling with anti-GOBP1, sometimes with anti-ABPX. S. basiconica in both sexes most commonly co-expressed GOBP1 and GOBP2, but other patterns were occasionally found, with some of them showing PBP expression, also in females. The great variety of labelling types in short s. trichodea, s. intermedia, and s. basiconica suggests a similar variety of functional subtypes as observed in plant odour-sensitive sensilla of other moth species.