| 日本血吸虫童虫表膜结合多肽的亲和筛选与初步鉴定 |
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| 引用本文： | 魏琦,刘彦,曾庆仁,陈欲晓,周军,刘碧源,邹艳,杨胜辉,蔡力汀,李立新,兰玲梅.日本血吸虫童虫表膜结合多肽的亲和筛选与初步鉴定[J].中国寄生虫学与寄生虫病杂志,2010,28(3):172-175. |
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| 作者姓名： | 魏琦 刘彦 曾庆仁 陈欲晓 周军 刘碧源 邹艳 杨胜辉 蔡力汀 李立新 兰玲梅 |
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| 作者单位： | 1 中南大学基础医学院免疫学教研室，长沙 410013；2 中南大学基础医学院细胞与分子生物学实验中心，长沙 410013；3 中南大学湘雅三医院实验中心，长沙 410013；4 湖南中医药高等专科学校基础医学部，株洲 410013 |
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| 基金项目： | 国家自然科学基金(No.30570952); 湖南省卫生厅(2009)项目~~ |
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| 摘 要： | 目的筛选噬菌体十二肽库中与日本血吸虫(Schistosoma japonicum)童虫表膜特异性结合的多肽并鉴定。方法利用噬菌体十二肽库与日本血吸虫活童虫靶分子之间的亲和力结合,经3轮吸附-洗脱-扩增,从末次回收的结合噬菌体中随机挑取20个克隆进行测序。根据测序结果 ,选取出现次数最多的噬菌体克隆作为目标噬菌体,免疫组化检测目的噬菌体,并回输到已感染日本血吸虫的小鼠体内,2.5h后处死小鼠,回收肝脏和日本血吸虫童虫,分别洗脱与肝脏和童虫结合的噬菌体,进行统计学分析。结果经3轮筛选后,噬菌体回收率从第1轮的0.77×10-8到第3轮的0.75×10-5,说明噬菌体得到了有效富集。DNA测序结果表明,20个噬菌体克隆中的15个克隆呈现QHPRIRKOOOOO序列。免疫组化结果显示,表达该序列的噬菌体能与童虫表膜有效结合;体内回输试验证实,该噬菌体能有效地靶向结合于体内日本血吸虫童虫表膜。结论筛选获得的短肽QHPRIRKOOOOO能有效地靶向结合日本血吸虫童虫表膜。
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| 关 键 词： | 日本血吸虫 童虫 噬菌体展示技术 表膜 靶向性 |
Screening and Characterization of Peptides Specifically Binding to the Schistosomulum Tegument of Schistosoma japonicum |
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| WEI Qi,LIU Yan,ZENG Qing-ren,CHEN Yu-xiao,ZHOU Jun,LIU Bi-,ZOU Yan,YANG Sheng-hui,CAI Li-ting,LI Li-xin,LAN Ling-mei.Screening and Characterization of Peptides Specifically Binding to the Schistosomulum Tegument of Schistosoma japonicum[J].Chinese Journal of Parasitology and Parasitic Diseases,2010,28(3):172-175. |
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| Authors: | WEI Qi LIU Yan ZENG Qing-ren CHEN Yu-xiao ZHOU Jun LIU Bi- ZOU Yan YANG Sheng-hui CAI Li-ting LI Li-xin LAN Ling-mei |
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| Institution: | WEI Qi1,2,LIU Yan2,ZENG Qing-ren2,CHEN Yu-xiao1,ZHOU Jun3,LIU Bi-yuan1,ZOU Yan4,YANG Sheng-hui2,CAI Li-ting2,LI Li-xin1,LAN Ling-mei2(1 Department of Immunology,Xiangya Medical School,the Central South University,Changsha 410013,China,2 Cell & Molecular Biological Experiment Xiang Ya School of Medicine Center,3 the Third Xiangya Hospital of Central South University,4 Hunan Traditional Chinese Medical College,Zh... |
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| Abstract: | Objective To screen and analyze the peptides in 12 phage-display peptide library specifically binding to the schistosomulum tegument of Schistosoma japonicum. Methods A 12 phage-display peptide library was screened with the S. japonicum schistosomula as the target cells for biopanning by degrees, positive clones picked randomly were deduced by DNA sequencing. According the sequencing result, immunohistochemical staining was performed to determine the specificity of the phages to the tegument. To test their ... |
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| Keywords: | Schistosoma japonicum Schistosomulum Phage display Tegument Target |
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