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| 免疫筛选旋毛虫cDNA克隆及原核表达 | |
| 引用本文： | 杨静,诸欣平,杨雅平,周蕾,Pascal Boireau,詹斌,冯建军,Hotez Peter.免疫筛选旋毛虫cDNA克隆及原核表达[J].中国病原生物学杂志,2003,16(1):7-9. |
| 作者姓名： | 杨静 诸欣平 杨雅平 周蕾 Pascal Boireau 詹斌 冯建军 Hotez Peter |
| 作者单位： | 1. 首都医科大学基础医学院寄生虫学教研室,北京 100054 2. UMR BIPAR INRA AFSSA ENVA UPVM, Maisons Alfort France 3. Department of Microbiology and Tropical medicine, George Washington University, Washington DC, USA, 20037 |
| 基金项目： | 美国中华医学会 (CMB) (Parasitology 98- 674)，国家留学基金回国科研资助 (1 998- 2 0 0 1 )，北京市教委科技发展基金 (2 0 0 1 - 2 0 0 4 )，北京市跨世纪优秀人材工程 (2 0 0 1 - 2 0 0 3)资助项 目。ING O |
| 摘 要： | 目的 获取旋毛虫抗原的 c DNA克隆并进行蛋白的原核表达。 方法 应用兔抗旋毛虫成虫可溶性全虫抗原血清对旋毛虫成虫 c DNA文库进行筛选 ,并用兔人工感染旋毛虫血清对强阳性克隆进行再筛选。将编号为 Ts87阳性克隆的基因片段亚克隆入 PET- 2 8a( +)表达载体 ,IPTG诱导表达后用 SDS- PAGE电泳分析表达产物。 结果 免疫筛选获得阳性克隆 Ts87;成功构建重组表达质粒 PET- 2 8a( +) / Ts87。诱导表达该融合蛋白 ,SDS- PAGE电泳表明 ,其能表达一分子质量约为 40 ku的融合蛋白 ,与预测分子质量相符。 结论 筛选到 c DNA克隆 Ts87,与兔抗旋毛虫成虫可溶性全虫抗原血清和兔人工感染旋毛虫血清均产生特异性免疫反应 ;PET原核表达系统所获重组蛋白为蛋白功能研究奠定了基础。 |
| 关 键 词： | 旋毛虫 免疫筛选 cDNA克隆 原核表达 |
| 文章编号： | 1001-6627(2003)01-0007-03 |
| 修稿时间： | 2002年3月14日 |
IMMUNOSCREENING OF TRICHINELLA SPIRALIS cDNA CLONES AND PROKARYOTIC EXPRESSION |
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| Pascal Boireau,Hotez Peter.IMMUNOSCREENING OF TRICHINELLA SPIRALIS cDNA CLONES AND PROKARYOTIC EXPRESSION[J].Journal of Pathogen Biology,2003,16(1):7-9. | |
| Authors: | Pascal Boireau Hotez Peter |
| Abstract: | Objective To obtain cDNA clones of Trichinella spiralis antigen and express protein. Methods An adult cDNA library of T. spiralis was screened by using rabbit immune sera, the strong positive clones were screened again by using T. spiralis-infected rabbit sera. The gene of positive clone Ts87 was subcloned into the expression vector PET-28a(+) to construct the expression plasmid. The transformants was induced by IPTG and its expression product analyzed by SDS-PAGE. Results Positive clones Ts87 was obtained, and recombinant plasmid PET-28a(+)/Ts87 was constructed successfully. The target recombinant proteins with a predicted relative molecular weight of 40 ku was got. Conclusion cDNA clone Ts87 was screened and reacted with rabbit immune sera and T. spiralis-infected rabbit sera. The target recombinant protein obtained by PET system laid a foundation of protein function research. |
| Keywords: | Trichinella spiralis immunoscreening cDNA clone prokaryotic expression |
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