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Babesia divergens: identification and characterization of BdHSP-20, a small heat shock protein.

机译:巴氏杆菌(Babesia divergens):一种小的热激蛋白BdHSP-20的鉴定和表征。

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This study describes the identification and characterization of the Babesia divergens-crystallin/small heat shock protein 20 (BdHSP-20). BdHSP-20 was recognized by the DG7 monoclonal antibody (DG7 mAb) originally produced by Precigout et al. [Precigout, E., Valentin, A., Carcy, B., Gorenflot, A., Nakamura, K., Aikawa, M., Schrevel, J. 1993. Babesia divergens: characterization of a 17-kDa merozoite membrane protein. Experimental Parasitology 77, 425-434] against B. divergens merozoites. We used DG7 mAb to immunoscreen a B. divergens cDNA library to clone the gene encoding the small heat shock protein. Bdhsp-20 is a single copy gene interrupted by one intron. The deduced gene product (BdHSP-20) clearly belongs to the alpha -crystallin family and shows significant homology to Babesia bovis, Plasmodium falciparum and Toxoplasma gondii sHSPs, with the highest degree of sequence identity around the catalytic domain. Nutrient stress (serum depletion) treatment of the parasites induced the upregulation of BdHSP-20 gene expression observed by semi-quantitative PCR and immunoprecipitation. This regulation pattern suggests that BdHSP-20 could probably be of importance for parasite survival in the case of environmental stress. BdHSP-20 has previously been shown to be highly conserved among different strains and antibodies against the protein drastically reduce parasitemia in vitro.
机译:这项研究描述了巴贝斯多巴菌晶体蛋白/小热激蛋白20(BdHSP-20)的鉴定和表征。 Predigout等人最初生产的DG7单克隆抗体(DG7 mAb)识别BdHSP-20。 [Precigout,E.,Valentin,A.,Carcy,B.,Gorenflot,A.,Nakamura,K.,Aikawa,M.,Schrevel,J. 1993.巴贝斯分枝杆菌:17 kDa裂殖子膜蛋白的表征。实验寄生虫学77,425-434]针对B. divergens裂殖子。我们使用DG7 mAb免疫筛选了B. divergens cDNA文库,以克隆编码小热激蛋白的基因。 Bdhsp-20是被一个内含子打断的单拷贝基因。推导的基因产物(BdHSP-20)显然属于α-晶状蛋白家族,并且与牛肝杆菌,恶性疟原虫和弓形虫sHSPs具有显着同源性,在催化域附近具有最高程度的序列同一性。对寄生虫的营养应激(血清耗竭)处理诱导了通过半定量PCR和免疫沉淀观察到的BdHS​​P-20基因表达的上调。这种调节模式表明,在环境压力的情况下,BdHSP-20可能对寄生虫的生存很重要。以前已显示BdHSP-20在不同菌株中是高度保守的,并且针对该蛋白的抗体可在体外显着降低寄生虫病。

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