日本血吸虫新抗原基因Sj-Ts4的克隆、表达及免疫保护性(英文)  

Cloning, Expression and Immunization of the New Antigen Gene Sj-Ts4 of Schistosoma japonicum

作　　者：周东明[1] 易新元[1] 曾宪芳[1] 王敏[1] 蔡春[1] 王庆林[1] 张顺科[1] MCREYNOLDS Larry 

机构地区：[1]中南大学湘雅医学院寄生虫学教研室,长沙410078 [2]Molecular Parasitology Group,New England Biolabs.Inc,ma01915

出　　处：《生物化学与生物物理学报》2002年第2期143-148,共6页

基　　金：WHO/TDR血吸虫病疫苗资助项目 (No .980 2 6 8)~~

摘　　要：为探讨旋毛虫感染小鼠后抗血吸虫感染的分子机制 ,并为血吸虫病疫苗的研究提供新的抗原分子 ,用旋毛虫感染鼠血清筛选日本血吸虫成虫cDNA文库。经过 3轮筛选 ,获得 9个阳性克隆 ,其中新基因Sj Ts4有一完整的编码框 ,编码2 10个氨基酸。该蛋白质的理论分子量为 2 3kD ,等电点为 7.72。将Sj Ts4亚克隆于原核表达载体 pGEX 5X 3,以GST融合蛋白的形式在E .coliDH5α中表达。Western印迹分析显示 ,重组Sj Ts4蛋白 (rSj Ts4)能被慢性感染鼠血清识别。rSj Ts4或与弗氏完全佐剂混合后免疫小鼠 ,均可以诱导产生特异性的IgG抗体 ,抗体效价可高达 1∶2 5 6 0 0。两免疫组的减虫率分别为31.36 %和 36 .80 % 。In order to explore the molecular mechanism of high immune protection against schistosomes infection in animals infected with Trichinella spiralis, and to provide several cross-protective antigen genes for developing anti-schistosomiasis vaccine, a Schistosoma japonicum adult worm cDNA library was immunoscreened using sera taken from mice infected with Trichinella spiralis. Nine positive clones were obtained after 3 rounds of immunoscreening. Among them, Sj-Ts4 represents a novel gene of S. japonicum, which coding for a novel protein of 210 amino acids. The protein has a deduced molecular mass of 23 kD and isoelectric point of 7.72. Sj-Ts4 was expressed as a glutathione-S-transferase (GST) fusion protein by cloning into the prokaryotic expression vector pGEX-5X-3. The recombinant Sj-Ts4 protein (rSj-Ts4) was purified and could be recognized by sera of mice infected with S.japonicum. Vaccination of several groups of mice with rSj-Ts4 or rSj-Ts4 incorporated into Freund's complete adjuvant induced high titers of specific IgG antibodies. Two vaccination groups all obtained significant reduction in worm burden ( 31.36%, 36.80%, P<0.01), compared with the control groups.

关 键 词：日本血吸虫 旋毛虫 Sj-Ts4 分子克隆 免疫保护性 抗原分子 基因表达 

分 类 号：Q78[生物学—分子生物学]