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多房棘球绦虫原头蚴脂肪酸结合蛋白基因的克隆与鉴定。

Cloning and characterization of the fatty acid-binding protein gene from the protoscolex of Taenia multiceps.

机构信息

College of Veterinary Medicine, Sichuan Agricultural University, Ya'an, 625014, China.

出版信息

Parasitol Res. 2013 May;112(5):1833-9. doi: 10.1007/s00436-013-3328-0. Epub 2013 Mar 9.

Abstract

Taenia multiceps (Cestoda: Taeniidae), a worldwide cestode parasite, is emerging as an important helminthic zoonosis due to serious or fatal central nervous system disease commonly known as coenurosis in domestic and wild ruminants including humans. Herein, a fatty acid-binding protein (FABP) gene was identified from transcriptomic data in T. multiceps. This gene, which contains a complete coding sequence, was amplified by reverse-transcriptase polymerase chain reaction. The corresponding protein, which was named TmFABP, had a molecular weight of 14 kDa, and subsequently was recombinantly expressed in Escherichia coli. The fusion protein was purified on Ni-NTA beads (Bio-Rad). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analyses showed that the purified recombinant protein caused immunogenicity. Immunohistochemical studies showed that TmFABP was expressed at the tegumental level in the protoscolices and in the cells between the body wall and parenchyma layer of the cestode. In sections from gravid proglottids, intense staining was detected in the uterus and eggs. Based on this, TmFABP could be switched on during differentiation of germinative layers to protoscoleces and from metacestodes to adult worms. Taken together, our results already reported for T. multiceps suggest the possibility of TmFABP developing a vaccine to control and prevent coenurosis.

摘要

多头绦虫(带绦虫科)是一种全球性的绦虫寄生虫,由于在包括人类在内的家养和野生反刍动物中常见的中枢神经系统疾病(通常称为多头蚴病),它正成为一种重要的寄生虫性人畜共患病。在此,从多头绦虫的转录组数据中鉴定出一种脂肪酸结合蛋白(FABP)基因。该基因包含一个完整的编码序列,通过逆转录聚合酶链反应扩增。相应的蛋白质被命名为 TmFABP,分子量为 14 kDa,随后在大肠杆菌中重组表达。融合蛋白通过 Ni-NTA 珠(Bio-Rad)进行纯化。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和 Western blot 分析表明,纯化的重组蛋白具有免疫原性。免疫组织化学研究表明,TmFABP 在原头节的表皮层和绦虫体壁与实质层之间的细胞中表达。在有孕节片中,在子宫和卵中检测到强烈的染色。基于此,TmFABP 可能在生殖层向原头节以及从囊尾蚴向成虫的分化过程中被激活。综上所述,我们已经报道的多头绦虫的结果表明,TmFABP 有可能开发成疫苗来控制和预防多头蚴病。

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