| 旋毛虫Ts87抗原基因在毕赤酵母中的构建及表达 |
| |
| 引用本文： | 王少华,诸欣平,顾园,杨雅平,杨静. 旋毛虫Ts87抗原基因在毕赤酵母中的构建及表达[J]. 中国寄生虫学与寄生虫病杂志, 2005, 23(4): 236-239 |
| |
| 作者姓名： | 王少华 诸欣平 顾园 杨雅平 杨静 |
| |
| 作者单位： | 首都医科大学基础医学院人体寄生虫学教研室,北京,100054 |
| |
| 基金项目： | 美国中华医学会基金（CMB）（Parasitology 98-674）;北京市教委科技发展基金（01KJ-091）;北京市跨世纪优秀人才工程（2001-2004）;首都医科大学校青年基金（2002-2005） |
| |
| 摘 要： | 目的构建旋毛虫Ts87抗原基因并在毕赤酵母中分泌表达。方法通过PCR特异性扩增Ts87抗原基因,构建重组质粒PPICZαA-Ts87。其线性化后,转化毕赤酵母GS115,抗生素Zeocin筛选高抗性转化子,PCR筛选阳性重组菌株。表型鉴定后进行甲醇诱导表达,收集培养不同天数的上清液。斑点杂交法分析上清液以确定有无重组蛋白分泌表达。十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质印迹法(Western blotting)分析鉴定表达蛋白。结果与结论成功构建了PPICZαA-Ts87毕赤酵母表达重组质粒,确定了Ts87抗原基因在毕赤酵母中获得分泌表达。
|
| 关 键 词： | 旋毛虫 蠕虫基因 毕赤酵母菌 基因表达 |
| 文章编号： | 1000-7423（2005）-04-0236-04 |
| 收稿时间： | 2004-09-17 |
| 修稿时间： | 2004-09-17 |
Construction and Expression of the Ts87 Gene of Trichinella spiralis in Pichia pastoris |
| |
| WANG Shao-hua,ZHU Xin-ping,GU Yuan,YANG ya-ping,YANG Jing. Construction and Expression of the Ts87 Gene of Trichinella spiralis in Pichia pastoris[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2005, 23(4): 236-239 |
| |
| Authors: | WANG Shao-hua ZHU Xin-ping GU Yuan YANG ya-ping YANG Jing |
| |
| Affiliation: | Department of Parasitology, School of Basic Medicine, Capital University of Medical Sciences, Beijing 100054, China. |
| |
| Abstract: | OBJECTIVE: To construct the recombinant plasmid P(PICZ) alphaA-Ts87 and obtain secretive protein in Pichia pastoris. METHODS: Ts87 was amplified by PCR and cloned into the expression vector P(PICZ) alphaA of P. pastoris to form P(PICZ) alphaA-Ts87. After the recombinant was isolated and linearized, it was transformed into P. pastoris GS115 by EasyComp kit and screened for zeocin resistant with different conditions and Mut phenotype. The high resistant Mut+ clones were cloned and the product induced by methanol was tested by dot hybridization. The expression product was further identified by SDS-PAGE and Western blotting. RESULTS AND CONCLUSION: The recombinant plasmid P(PiCZ) alphaA-Ts87 has been constructed. Ts87 gene has been expressed and secreted in Pichia pastoris GS115 strain. |
| |
| Keywords: | Trichinella spiralis Helminth genes Pichia pastoris Gene expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
| 点击此处可从《中国寄生虫学与寄生虫病杂志》浏览原始摘要信息 |
|
点击此处可从《中国寄生虫学与寄生虫病杂志》下载免费的PDF全文 |
