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鉴定和特性分析肌联蛋白样蛋白:华支睾吸虫的生活史阶段和组织特异性抗原。

Identification and characterization of myophilin-like protein: a life stage and tissue-specific antigen of Clonorchis sinensis.

机构信息

Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, China.

出版信息

Parasitol Res. 2012 Sep;111(3):1143-50. doi: 10.1007/s00436-012-2946-2. Epub 2012 Jun 30.

Abstract

Clonorchiasis, caused by Clonorchis sinensis infection, has been an important public health problem in China. More messages about biology and pathogenicity of C. sinensis will be better for development of new strategies for clonorchiasis control. In the current study, using data from the published genomic and cDNA sequences for C. sinensis, we identify a gene encoding a myophilin-like protein (Cs myophilin-like). The cDNA sequence is 573 bp in length which encoded a protein of 190 amino acids. DNA sequence of Cs myophilin-like consists of four extrons and three introns. By mapping the draft genome of C. sinensis, single copy of myophilin-like gene is annotated. Bioinformatic analysis shows Cs myophilin-like comprises a calponin homology domain and three actin binding surfaces. It expresses at the life stage of adult worm and metacercaria but not in eggs. It intensively distributes in muscular locations such as oral sucker and pharynx of adult worm and sucker of metacercaria. The transcript level of Cs myophilin-like in oral sucker and pharynx is significantly higher than that in the rest of adult worm. Moreover, recombinant Cs myophilin-like is obtained and of antigenicity and immunoreactivity. It can be probed by sera from patients infected with trematode or cestode. Our data suggests that Cs myophilin-like may be involved in the host tissue invasion besides regulating the contraction of smooth muscle and movement of worm body. Taken together, the stable and soluble expression in Escherichia coli and immunogenicity of recombinant Cs myophilin-like proteins suggest that Cs myophilin-like may be a potential candidate for vaccine and drug target for clonorchiasis.

摘要

华支睾吸虫病是由华支睾吸虫感染引起的,一直是中国重要的公共卫生问题。更多关于华支睾吸虫的生物学和致病性的信息将有助于开发新的华支睾吸虫病控制策略。在本研究中,我们利用已发表的华支睾吸虫基因组和 cDNA 序列数据,鉴定了一个编码肌球蛋白轻链样蛋白(Cs 肌球蛋白轻链样)的基因。cDNA 序列长 573bp,编码 190 个氨基酸的蛋白质。Cs 肌球蛋白轻链样的 DNA 序列由四个外显子和三个内含子组成。通过对华支睾吸虫基因组草图进行映射,鉴定出一个肌球蛋白轻链样基因的单拷贝。生物信息学分析表明,Cs 肌球蛋白轻链样包含一个钙调蛋白同源结构域和三个肌动蛋白结合面。它在成虫和囊蚴的生活阶段表达,但不在卵中表达。它在成虫的口吸盘和咽以及囊蚴的吸盘等肌肉部位高度分布。在口吸盘和咽中 Cs 肌球蛋白轻链样的转录水平显著高于成虫的其他部位。此外,还获得了重组 Cs 肌球蛋白轻链样蛋白,并具有抗原性和免疫反应性。它可以被感染吸虫或绦虫的患者血清所探测。我们的数据表明,Cs 肌球蛋白轻链样可能除了调节平滑肌收缩和虫体运动外,还可能参与宿主组织侵袭。综上所述,重组 Cs 肌球蛋白轻链样蛋白在大肠杆菌中的稳定和可溶性表达以及免疫原性表明,Cs 肌球蛋白轻链样可能是华支睾吸虫病疫苗和药物靶点的潜在候选物。

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