Molecular cloning and immunological characterization of phosphoglycerate kinase from Clonorchis sinensis
Molecular cloning and immunological characterization of phosphoglycerate kinase from Clonorchis sinensis
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DOI：
10.1016/s0166-6851(00)00220-6
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发表时间：
2000-05-01
影响因子：
1.5
通讯作者：
Song, KY
中科院分区：
文献类型：
--
作者：
Hong, SJ;Seong, KY;Song, KY
寄生蜂华支睾吸虫（Clonediumsinensis）是一种利用大量外界葡萄糖进行能量代谢的寄生虫。磷酸甘油酸激酶（PGK）是一种存在于寄生虫体内的糖酵解酶，被认为是疫苗和药物开发的候选分子之一。通过筛选C.利用异源cDNA探针构建的中华猪PGKs cDNA文库编码415个氨基酸，与多种动物PGKs的同源性超过60%。推定的肽揭示了对应于12个β-折叠的结构域和形成动物PGKs的底物结合裂缝的内环。该基因产物在大肠杆菌中过表达，并显示PGK样酶活性。制备了抗重组C. sinensis PGK对天然C.结果表明，PGK在中华绒螯蟹成虫的肌肉组织和体壁中均有分布。梭PGK在C. sinensis中诱导产生抗体。感染了中华按蚊的兔子因此，建议将C.结论：华支睾吸虫PGK可作为华支睾吸虫病血清学诊断的免疫试剂。(C)2000 Elsevier Science B. V.保留所有权利。
The parasite Clonorchis sinensis was determined to utilize a large amount of external glucose to carry its energy metabolism. Phosphoglycerate kinase (PGK), a glycolytic enzyme, found in many parasites, has been identified as one of the candidate molecules distinguished from human counterparts for vaccine and drug developments. A cDNA clone purified by screening a C. sinensis cDNA library using a heterologous cDNA probe encoded a putative peptide of 415 amino acids with over 60% identities with PGKs from a number of animals. The putative peptides revealed domains corresponding to 12 beta-sheets and inner loops forming a substrate-binding cleft of animal PGKs. The gene product was overexpressed in Escherichia coli and showed a PGK-like enzyme activity. A polyclonal antibody raised against the recombinant C. sinensis PGK was specific to native C. sinensis PGK and localized it to the muscular tissue and tegument of the adult flukes. The C. sinensis PGK elicited antibodies in C. sinensis-infected rabbits. Therefore, it is proposed that C. sinensis PGK could be used as an immunoreagent in the serodiagnosis for clonorchiasis. (C) 2000 Elsevier Science B.V. All rights reserved.