Guan Guiquan, Liu Junlong, Liu Aihong, Li Youquan, Niu Qingli, Gao Jinliang, Luo Jianxun, Chauvin Alain, Yin Hong, Moreau Emmanuelle
State Key Laboratory of Veterinary Etiological Biology,Key Laboratory of Veterinary Parasitology of Gansu Province,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Science,Xujiaping 1,Lanzhou,Gansu 730046,People's Republic of China.
LUNAM Université,Oniris,Ecole nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique,UMR Biologie,Epidémiologie et Analyse de Risque en santé animale,CS 40706,F-44307 Nantes,France.
Parasitology. 2015 Sep;142(11):1387-97. doi: 10.1017/S0031182015000797. Epub 2015 Jul 9.
Heat shock protein 90 (HSP90) is a key component of the molecular chaperone complex essential for activating many signalling proteins involved in the development and progression of pathogenic cellular transformation. A Hsp90 gene (BQHsp90) was cloned and characterized from Babesia sp. BQ1 (Lintan), an ovine Babesia isolate belonging to Babesia motasi-like group, by screening a cDNA expression library and performing rapid amplification of cDNA ends. The full-length cDNA of BQHsp90 is 2399 bp with an open reading frame of 2154 bp encoding a predicted 83 kDa polypeptide with 717 amino acid residues. It shows significant homology and similar structural characteristics to Hsp90 of other apicomplex organisms. Phylogenetic analysis, based on the HSP90 amino acid sequences, showed that the Babesia genus is clearly separated from other apicomplexa genera. Five Chinese ovine Babesia isolates were divided into 2 phylogenetic clusters, namely Babesia sp. Xinjiang (previously designated a new species) cluster and B. motasi-like cluster which could be further divided into 2 subclusters (Babesia sp. BQ1 (Lintan)/Babesia sp. Tianzhu and Babesia sp. BQ1 (Ningxian)/Babesia sp. Hebei). Finally, the antigenicity of rBQHSP90 protein from prokaryotic expression was also evaluated using western blot and enzyme-linked immunosorbent assay (ELISA).
热休克蛋白90(HSP90)是分子伴侣复合物的关键组成部分,对于激活许多参与致病性细胞转化发展和进程的信号蛋白至关重要。通过筛选cDNA表达文库并进行cDNA末端快速扩增,从属于莫氏巴贝斯虫样组的绵羊巴贝斯虫分离株——巴贝斯虫sp. BQ1(临潭)中克隆并鉴定了一个Hsp90基因(BQHsp90)。BQHsp90的全长cDNA为2399 bp,开放阅读框为2154 bp,编码一个预测的83 kDa多肽,含有717个氨基酸残基。它与其他顶复门生物的Hsp90具有显著的同源性和相似的结构特征。基于HSP90氨基酸序列的系统发育分析表明,巴贝斯虫属与其他顶复门属明显分开。五个中国绵羊巴贝斯虫分离株分为2个系统发育簇,即新疆巴贝斯虫(以前被指定为一个新物种)簇和莫氏巴贝斯虫样簇,后者可进一步分为2个亚簇(巴贝斯虫sp. BQ1(临潭)/巴贝斯虫sp. 天祝和巴贝斯虫sp. BQ1(宁县)/巴贝斯虫sp. 河北)。最后,还使用蛋白质印迹法和酶联免疫吸附测定(ELISA)评估了原核表达的rBQHSP90蛋白的抗原性。