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微小扇头蜱(牛蜱属):Bm86-CG在毕赤酵母中的表达及特性分析

Rhipicephalus (Boophilus) microplus: expression and characterization of Bm86-CG in Pichia pastoris.

作者信息

Cunha Rodrigo Casquero, Andreotti Renato, Leite Fábio Pereira Leivas

机构信息

Universidade Federal de Mato Grosso do Sul, Campo Grande, MS, Brazil.

出版信息

Rev Bras Parasitol Vet. 2011 Apr-Jun;20(2):103-10. doi: 10.1590/s1984-29612011000200003.

Abstract

The cattle tick Rhipicephalus (Boophilus) microplus is responsible for great economic losses. It is mainly controlled chemically, with limitations regarding development of resistance to the chemicals. Vaccines may help control this parasite, thereby reducing tick pesticide use. In this light, we performed subcloning of the gene of the protein Bm86-GC, the homologue protein that currently forms the basis of vaccines (Gavac(TM) and TickGard(PLUS)) that have been developed against cattle ticks. The subcloning was done in the pPIC9 expression vector, for transformation in the yeast Pichia pastoris. This protein was characterized by expression of the recombinant Mut+ strain, which expressed greater quantities of protein. The expressed protein (rBm86-CG) was recognized in the Western-blot assay using anti-Gavac, anti-TickGard, anti-larval extract and anti-rBm86-CG polyclonal sera. The serum produced in cattle vaccinated with the antigen CG rBm86 presented high antibody titers and recognized the native protein. The rBm86-GC has potential relevance as an immunogen for vaccine formulation against cattle ticks.

摘要

牛蜱微小牛蜱(Rhipicephalus (Boophilus) microplus)会造成巨大的经济损失。其主要通过化学方法进行防治,但在化学药物抗性发展方面存在局限性。疫苗可能有助于控制这种寄生虫,从而减少蜱虫杀虫剂的使用。有鉴于此,我们对蛋白质Bm86 - GC的基因进行了亚克隆,该蛋白质是目前已开发的针对牛蜱的疫苗(Gavac™和TickGard(PLUS))的同源蛋白。亚克隆是在pPIC9表达载体中进行的,用于在毕赤酵母(Pichia pastoris)中转化。该蛋白质通过重组Mut +菌株的表达进行表征,该菌株表达了更多数量的蛋白质。使用抗Gavac、抗TickGard、抗幼虫提取物和抗rBm86 - CG多克隆血清在蛋白质免疫印迹分析中识别出表达的蛋白质(rBm86 - CG)。用抗原CG rBm86免疫的牛产生的血清呈现出高抗体滴度并识别天然蛋白质。rBm86 - GC作为针对牛蜱的疫苗制剂的免疫原具有潜在的相关性。

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