新型pⅨ噬菌体展示系统的建立及初步评价  

Construction of a new phagemid for phage display using phage coat protein pⅨ and its primary evaluation

作　　者：王双[1] 孙志伟[1] 杜威世[1] 张锦超[1] 俞炜源[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《军事医学科学院院刊》2005年第5期444-446,共3页Bulletin of the Academy of Military Medical Sciences

基　　金：国家"973"课题(2002CB513205)

摘　　要：目的:构建新型噬菌体展示载体,评价其展示效果,为构建scFv库寻找新的展示体系。方法:PCR分离、扩增M13KO7中pⅨ蛋白编码基因,克隆入噬菌体展示载体PHB-gⅢ,替换原载体中的gⅢ-CT段,构建载体PHB-pⅨ,并将相对分子质量约50×103的碱性磷酸酶的编码基因分别克隆入两展示载体,观察其展示碱性磷酸酶能力的差别。结果与结论:成功构建了展示载体PHB-pⅨ,该载体具有与PHB-gⅢ相当的展示碱性磷酸酶的能力,且相同滴度的PHB-gⅢ噬菌体DNA含量略高于PHB-pⅨ噬菌体DNA含量,提示PHB-gⅢ系统展示外源蛋白时存在感染能力下降。PHB-pⅨ系统具有用于展示噬菌体抗体库的良好应用前景。Objective： To construct a new phage display vector named PHB-pⅨ and evaluate its display ability in order to find a novel display system for antibody library. Methods： pⅨ gene was obtained from M13KO7 using PCR and was cloned into vector PHB-gⅢ to replace the gⅢ-CT, and the vector was named PHB-pⅨ. The display ability was compared between two systems by displaying alkaline phosphatase（Ap） with molecular weight about 50 × 10^3. Results and conclusion： Expression vector PHB-plX had equivalent-ability in displaying Ap, but PCR experiment indicated that the phagmid DNA number of PHB-gⅢ was higher than PHB-pⅨ in the same colony forming unit（cfu） , which suggested a reduced infection ability of PHB-gⅢ when displayed large proteins. It seemed that PHB-pⅨ will be a good phage display vector for antibody libraries.

关 键 词：噬菌体展示 pⅨ gⅢ 抗体库 

分 类 号：R346[医药卫生—基础医学]