大肠杆菌表面 CS3菌毛展示随机肽库的构建
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刘向昕, 袁盛凌, 展德文,郑继平, 刘纯杰, 王
CS3菌毛;细菌表面展示;随机肽库刘向昕,袁盛凌,展德文,郑继平,刘纯杰,王芃,王令春,张兆山.大肠杆菌表面CS3菌毛展示随机肽库的构建.世界华人消化杂志2006;14(2)158-162,大肠杆菌表
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刘向昕, 袁盛凌,展德文,郑继平,刘纯杰,王芃,王令春,张兆山,中国人民解放军军事医学科学院生物工程研究所 北京市 100071
刘向昕,女, 1974-07-07生,山西省太原市人汉族, 2004年中国人民解放军军事医学科学院博士生, 主要从事基因工程疫苗研究.
国家高技术研究发展863计划项目, No.2004AA215212
通讯作者:张兆山, 100071, 北京丰台区东大街20号,中国人民解放军军事医学科学院生物工程研究所.zhangzs@nic.bmi.ac.cn
电话: 010-63834140 传真: 010-63833521
收稿日期: 2005-11-08 接受日期: 2005-12-08
Construction of a CS3 fimbria-displayed randompeptide library on Escherichia coli cell surface
Xiang-Xin Liu, Sheng-Ling Yuan, De-WenZhan, Ji-Ping Zheng, Chun-Jie Liu, Peng Wang, Ling-Chun Wang, Zhao-ShanZhang
Xiang-Xin Liu, Sheng-Ling Yuan,De-Wen Zhan, Ji-Ping Zheng, Chun-Jie Liu, Peng Wang, Ling-Chun Wang,Zhao-Shan Zhang, Instituteof Biotechnology, Academy of Military Medical Sciences, Beijing 100071,China
Supported by theNational High Technology Research and Development Program of China (863Program), No.2004AA215212
Correspondence to: Zhao-ShanZhang, Institute of Biotechnology, Academy of Military Medical Sciences,20 Dongdajie Street, Fengtai District, Beijing 100071,China. zhangzs@nic.bmi.ac.cn
Received: 2005-11-08 Accepted: 2005-12-08
Abstract
AIM:To construct a CS3 fimbria-displayed random peptide library on the Escherichia coli cell surface
METHODS:Firstly, a new display vector with double restriction sites was reconstructed, and the ability of the new vector to form CS3 fimbriae on E. coli surface was confirmed by Western blot and transmission electron microscopy. Then two oligonucleotides were synthesized, one of which contained a random oligonucleotide encoding region (NNK)10 and the other was its complement. The two synthesized oligonucleotides was annealed and then extended by Klenow Fragment. The double-stranded oligonucleotides were digested by XhoT and BamHT and purified by PAGE, then inserted into the digested display vector. The ligation product was purified and electroporated into XL1-Blue ......
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