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利用噬菌体表面展示技术克隆抗人DR5抗体可变区基因
引用本文:刘瑞敏,王明丽,季泽俊,刘峰涛,白慧玲,马远方.利用噬菌体表面展示技术克隆抗人DR5抗体可变区基因[J].河南大学学报(医学版),2011,30(1):51-53.
作者姓名:刘瑞敏  王明丽  季泽俊  刘峰涛  白慧玲  马远方
作者单位:河南大学细胞与免疫学重点实验室,河南,开封,475004
基金项目:传染病预防控制国家重点实验室开放课题,河南省重点科技攻关资助项目
摘    要:目的:利用噬菌体表面展示技术制备抗人DR5单链抗体.方法:从抗DR5杂交瘤细胞中提取总RNA,RT-PCR扩增VH和VL基因,并利用重叠扩增PCR将VH和VL拼接为scFv基因.将scFv与PAK100载体通过相同的酶切位点连接后,转化大肠杆菌XL1-Blue,经VSCM13辅助噬菌体拯救,构建成鼠抗人DR5单链抗体库...

关 键 词:噬菌体展示技术  单链抗体  死亡受体5

Clone of the anti-DR5 antibody variable sequence by phage surface display
LIU Rui-min,WANG Ming-li,JI Ze-jun,LIU Feng-tao,BAI Hui-ling,Ma Yuan-fang.Clone of the anti-DR5 antibody variable sequence by phage surface display[J].Journal of Henan University,2011,30(1):51-53.
Authors:LIU Rui-min  WANG Ming-li  JI Ze-jun  LIU Feng-tao  BAI Hui-ling  Ma Yuan-fang
Institution:LIU Rui-min,WANG Ming-li,JI Ze-jun,LIU Feng-tao,BAI Hui-ling,Ma Yuan-fang(Key Laboratory of Cellular and Molecular Immunology,Henan University,Kaifeng,Henan 475000,China)
Abstract:Objective:To obtain the gene sequence of antibody against human death receptor 5 by the phage surface display.Methods:By the technique of phage display,the heavy chain and light chain variable region genes(VH and VL) were assembled into single chain antibodies(scFv) with overlap extension reaction by L inker primer mix.The scFv were cloned into PAK100 phagemid and introduced into E.coli XL1-Blue.The phagemid containing bacterial colonies were infected with VSCM13,then the total surface display library of an...
Keywords:Phage surface display technique  Single chain antibodies  Death receptor 5  
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