| 鼠源单抗3G1单链抗体基因的构建及序列分析 |
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| 引用本文： | 罗雯,阎岩,徐志凯,张芳琳,吴兴安,刘勇,白文涛,王海涛.鼠源单抗3G1单链抗体基因的构建及序列分析[J].西安联合大学学报,2003,6(2):5-8. |
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| 作者姓名： | 罗雯 阎岩 徐志凯 张芳琳 吴兴安 刘勇 白文涛 王海涛 |
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| 作者单位： | [1]西安联合大学化学与生命科学系，陕西西安710065 [2]第四军医大学微生物学教研室，陕西西安710032 |
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| 基金项目： | 国家自然科学基金资助项目 (3970 0 134) |
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| 摘 要： | 利用Rr—PCR方法，从分泌抗汉滩病毒mAb的杂交瘤细胞系3Gl中扩增出抗体VH和VL基因，通过PCR方法获得1inker—VL基因，进而利用加端PCR技术，在扩增出的单抗体VH和1inker—VL基因两端加上限制性酶切位点，分别克隆入pUCl8中并测序．3Gl VH基因长360bp，编码120个氨基酸；VL基因长324bp，编码108个氨基酸．在pUCl8载体中将VH和1inker—VL基因连接成ScFv基因．
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| 关 键 词： | 肾综合征出血热 鼠源单抗3Gl单链抗体基因 基因构建 序列分析 抗汉滩病毒 基因工程 |
| 文章编号： | 1008-777X(2003)02-0005-04 |
| 修稿时间： | 2002年6月8日 |
Construction and Sequence Analysis of the Single Chain Fv of Mouse Monoclonal Antibody 3G1 |
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| Authors: | LUO Wen YAN Yan XU Zhi kai ZHANG Fang lin WU Xing an LIU Yong BAI Wen tao WANG Hai tao |
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| Institution: | LUO Wen 1,YAN Yan 2,XU Zhi kai 2,ZHANG Fang lin 2,WU Xing an 2,LIU Yong 2,BAI Wen tao 2,WANG Hai tao 2 |
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| Abstract: | Total cell RNA was extracted from a hybridoma cell line 3G1 producing mouse mAb against HFRS virus, and the V H and V L genes were amplified by RT PCR. Linker V L gene was obtained by PCR, then the restriction enzyme sites were added on the previously amplified by add on PCR. The V H and Linker V L genes were respectively cloned into the vector pUC18 and sequenced. 3G1 V H gene consisted of 360bp, encoding 120 amino acids; V L gene consisted of 324 bp, encoding 108 amino acids. Linker V L and V H gene were joined to construct ScFv gene in pUC18 vector. |
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| Keywords: | Single Chain FV Gene Cloning Hemorrhagic Fever Virus Epidemic Antibody Monoclonal |
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