Immunoanalysis of unique protein in Trichoplusia ni larvae parasitized by the braconid wasp Chelonus near curvimaculatus

Ana I. Soldevila , Davy Jones
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引用次数: 17

Abstract

Parasitization in insects brings about profound biochemical and physiological effects in the host which may include complete overriding of the normal endocrinological program, resulting in precocious metamorphosis and in blockage of pupal development. The subtle effects of parasitization include changes in the expression of hemolymph proteins and the appearance of proteins which are unique to parasitized hosts. One such protein has been identified in the hemolymph of Trichoplusia ni larvae parasitized by the braconid wasp Chelonus near curvimaculatus. In this study, purified preparations of the parasitism-specific protein were used to generate polyconal antibodies against the protein. Results from the immunocharacterization indicate the antibodies obtained are highly specific for the protein and are present in a high titer (1:8000 antiserum dilution yielded strong signals in analysis of the protein in 0.25 μl hemolymph). Subsequently, the expression of the parasitism-specific protein in the hemolymph and tissues was analyzed by immunoblotting during the entire course of development in normal and parasitized insects. The parasitism-specific protein was not detected in normal, unparasitized larvae. In parasitized insects, expression of the parasitism-specific protein appears to be stage-specific in that it is only detected during the last larval stadium of precociously metamorphosing larvae, but is absent from all earlier stages of development.

屈斑蝶寄生蜂寄生的ni毛虫幼虫独特蛋白的免疫分析
寄生在昆虫体内会对寄主产生深远的生化和生理影响,包括完全改变正常的内分泌程序,导致早熟变态和蛹发育受阻。寄生的微妙影响包括血淋巴蛋白表达的变化和被寄生宿主特有的蛋白质外观。一种这样的蛋白已被发现存在于弯曲附近的斑蜂(Chelonus bronid wasp)寄生的ni毛虫幼虫的血淋巴中。在本研究中,利用纯化的寄生虫特异性蛋白制备了针对该蛋白的多孔抗体。免疫鉴定结果表明,获得的抗体对该蛋白具有高度特异性,并以高滴度存在(1:800抗血清稀释在0.25 μl血淋巴中分析该蛋白时产生强信号)。随后,利用免疫印迹法分析了正常和被寄生昆虫整个发育过程中血淋巴和组织中寄生特异性蛋白的表达。正常、未被寄生的幼虫中未检测到寄生特异性蛋白。在被寄生的昆虫中,寄生特异性蛋白的表达似乎是阶段性的,因为它只在早熟变态幼虫的最后一个幼虫场中被检测到,而在所有早期发育阶段都不存在。
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