| 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： | 2002-06-08 |
Construction and Sequence Analysis of the Single Chain Fv of Mouse Monoclonal Antibody 3G1 |
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| LUO Wen ,YAN Yan ,XU Zhi kai ,ZHANG Fang lin ,WU Xing an ,LIU Yong ,BAI Wen tao ,WANG Hai tao. Construction and Sequence Analysis of the Single Chain Fv of Mouse Monoclonal Antibody 3G1[J]. , 2003, 6(2): 5-8 |
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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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| Affiliation: | 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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