| 抗甲氧西林耐药金黄色葡萄球菌鼠源化噬菌体抗体库的构建及筛选 |
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| 引用本文： | 何光志,田维毅,高英,王平,王文佳,王乾宇,奚锦,俞琦,黄高,安传伟.抗甲氧西林耐药金黄色葡萄球菌鼠源化噬菌体抗体库的构建及筛选[J].广东农业科学,2011,38(13). |
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| 作者姓名： | 何光志 田维毅 高英 王平 王文佳 王乾宇 奚锦 俞琦 黄高 安传伟 |
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| 作者单位： | 1. 贵阳中医学院基础部,贵州,贵阳,550002
2. 遵义医学院附属医院,贵州,遵义,563003 |
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| 基金项目： | 贵阳中医学院博士基金; 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) |
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| 摘 要： | 用灭活的甲氧西林耐药金黄色葡萄球菌免疫小鼠后,取出小鼠脾脏提取总RNA,以RT-PCR扩增小鼠抗体重链(VH)和轻链(VL)可变区基因,以重叠延伸PCR(SOE-PCR)法将VH和VL拼接成ScFv基因,克隆入噬菌粒载体pCANTAB-5E中,转化大肠杆菌E.coli TG1,构建鼠源抗金黄色葡萄天然噬菌体抗体库。研究结果表明,构建的鼠源性抗甲氧西林耐药金黄色葡萄球菌抗体库库容为3.2×106,多样性良好,共筛选出8个阳性克隆。说明已成功构建抗甲氧西林耐药金黄色葡萄球菌抗体库,为筛选有效治疗甲氧西林耐药金黄色葡萄球菌奠定了基础。
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| 关 键 词： | 甲氧西林耐药金黄色葡萄球菌 抗体库 单链抗体库 |
Construction and screening of mice single-chain antibody library of anti-methicillin-resistant Staphylococcus aureus |
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| HE Guang-zhi,TIAN Wei-yi,GAO Ying,WANG Ping,WANG Wen-jia,WANG Qian-yu,XI Jin,YU Qi,HUANG Gao,AN Chuan-wei.Construction and screening of mice single-chain antibody library of anti-methicillin-resistant Staphylococcus aureus[J].Guangdong Agricultural Sciences,2011,38(13). |
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| Authors: | HE Guang-zhi TIAN Wei-yi GAO Ying WANG Ping WANG Wen-jia WANG Qian-yu XI Jin YU Qi HUANG Gao AN Chuan-wei |
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| Institution: | HE Guang-zhi1,TIAN Wei-yi1,GAO Ying2,WANG Ping1,WANG Wen-jia1,WANG Qian-yu1,XI Jin1,YU Qi1,HUANG Gao1,AN Chuan-wei1(1.Foundation Department,Guiyang college of Traditional Chinese Medicine,Guiyang 550002,China,2.Affiliated Hospital of Zunyi Medical College,Zunyi 563003,China) |
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| Abstract: | Mice were injected with anti-methicillin-resistant Staphylococcus aureus.Total RNA was extracted from the spleen cell of the immune mice followed RT-PCR.The heavy-chain and light-chain variable region genes(VH and VL) repertoire of immunoglobulin were amplified respectively by PCR.Then they were linked by a DNA linker encoding(G1y4 Ser) 3 as VH-linker-VL sequence forming scFv by SOE(splicing by overlap extension) PCR.These fragments were cloned into the phagemid pCANTAB5E and the recombinant phagemids were ... |
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| Keywords: | methicillin-resistant Staphylococcus aureus antibody library single-chain antibody |
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