Molecular cloning and characterization of heat shock protein 70 from Trichinella spiralis
Molecular cloning and characterization of heat shock protein 70 from Trichinella spiralis
复制标题
旋毛虫热休克蛋白 70 的分子克隆和表征。
DOI：
10.1016/j.actatropica.2009.01.003
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发表时间：
2009-04-01
期刊：
影响因子：
2.7
通讯作者：
Cui, Shijuan
中科院分区：
文献类型：
--
作者：
Wang, Shaohua;Zhu, Xinping;Cui, Shijuan
通过对旋毛虫成虫进行免疫筛选，获得了旋毛虫热休克蛋白70（Ts-Hsp 70）的cDNA序列。用兔抗旋毛虫血清构建旋毛虫cDNA文库。旋毛虫成虫提取物。Ts-Hsp 70（DNA）的开放阅读框编码623个氨基酸，预测分子量为68.7 kDa，与其他寄生虫的HSP 70具有高度的序列保守性。重组Ts-Hsp 70在大肠杆菌中表达，镍柱层析纯化。Westernblot分析表明，重组Ts-Hsp 70不仅能被旋毛虫病患者血清识别，而且能被旋毛虫感染的来自兔、猪和小鼠的螺旋体感染血清。用弗氏佐剂配制的重组Ts-Hsp 70免疫小鼠，产生了高滴度的IgG抗体，并在T.螺旋幼虫本研究结果表明，Ts-Hsp 70是一种可能的T.螺旋体感染(c)2009 Elsevier B. V.保留所有权利。
A cDNA encoding heat shock protein 70 of Trichinella spiralis (Ts-Hsp70) was identified by immunoscreening the adult T. spiralis cDNA library with rabbit antisera against T. spiralis adult extracts. The open reading frame of Ts-Hsp70 (DNA encoded a 623-amino acid peptide with a predicted molecular weight of 68.7 kDa, which shares a high degree of sequence conservation with HSP70s from other parasites. Recombinant Ts-Hsp70 was expressed in Escherichia coli and purified with nickel column chromatography. Western blot analysis showed that recombinant Ts-Hsp70 could be recognized not only by trichinellosis patient sera, but also by T. spiralis-infected sera from rabbits, swine, and mice. Mice vaccinated with recombinant Ts-Hsp70 formulated with Freund's adjuvant exhibited strong humoral immune responses indicated by high titer of IgG antibody and significant muscle larval reduction (37%) after being challenged with T. spiralis larvae. The present results indicate that Ts-Hsp70 is a possible candidate vaccine against T. spiralis infection. (c) 2009 Elsevier B.V. All rights reserved.