| 抗弓形虫噬菌体单链抗体库的构建及筛选 |
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| 引用本文： | 何光志,田维毅,王平,王文佳,奚锦,俞琦,黄高,王乾宇.抗弓形虫噬菌体单链抗体库的构建及筛选[J].湖北大学学报(自然科学版),2011,33(3):304-308. |
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| 作者姓名： | 何光志 田维毅 王平 王文佳 奚锦 俞琦 黄高 王乾宇 |
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| 作者单位： | 贵阳中医学院基础部,贵州贵阳,550002 |
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| 基金项目： | 贵阳中医学院博士基金; 贵州省卫生厅课题; 贵州省优秀科技教育人才省长资金项目(黔省专合字[2010]50号)资助 |
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| 摘 要： | 采用弓形虫可溶性抗原攻击的小鼠,取小鼠脾脏从中提取细胞总RNA,通过RT-PCR扩增鼠源抗体VH和VL基因,并采用重叠PCR (SOE-PCR)方法构建ScFv基因,将其克隆入噬粒载体pCANTAB5E中,转化于感受态大肠杆菌TG1,通过辅助噬菌体M13K07援救构建噬菌体单链抗体库.从20个噬菌体克隆中筛选到15个具...
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| 关 键 词： | 弓形虫 人源噬菌体单链抗体 制备及筛选 |
Construction and screening of single-chain antibody library of Toxoplasma gondii |
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| HE Guangzhi,TIAN Weiyi,WANG Ping,WANG Wenjia,XI Jin,YU Qi,HUANG Gao,WANG Qianyu.Construction and screening of single-chain antibody library of Toxoplasma gondii[J].Journal of Hubei University(Natural Science Edition),2011,33(3):304-308. |
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| Authors: | HE Guangzhi TIAN Weiyi WANG Ping WANG Wenjia XI Jin YU Qi HUANG Gao WANG Qianyu |
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| Institution: | HE Guangzhi,TIAN Weiyi,WANG Ping,WANG Wenjia,XI Jin,YU Qi,HUANG Gao,WANG Qianyu,(Department of Basic Courses,Guiyang College of Traditional Chinese Medicine,Guiyang 550002,China) |
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| Abstract: | A few of mice were injected Toxoplasma gondii RH,their spleen were removed and minced with a surgical knife,total RNA was extracted.To mice immunoglobulin H chain and L chain variable region of degenerate primers,cDNA first strand as a template,were amplified H chain and L chain variable region genes.SOE-PCR method to VH and VL fragments were assembled into ScFv fragments and then cloned into pCANTAB5E ScFv vector and transformed into competent TG1 by roporation.After rescuing with helper phage M13 K07.Fift... |
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| Keywords: | Toxoplasma gondii single-chain antibody construction and screening |
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