电转化技术在噬菌体抗体库构建中的应用  

The application of high-voltage gene pulser in constructing phage antibody repertoires

作　　者：赵禹[1] 侯秉璋[1] 乔玲[1] 刘嘉玉[1] 张墨玲[1] 李颖利[2] 陈昆明[1] 

机构地区：[1]天津医科大学内分泌研究所,天津300070 [2]天津医科大学总医院

出　　处：《天津医科大学学报》2002年第4期413-415,共3页Journal of Tianjin Medical University

基　　金：国家自然科学基金资助项目一部分 (批准号:39870336)

摘　　要：目的 :探讨电转化技术在构建噬菌体抗体库中的应用价值。方法 :用国产电转化仪将重组噬菌粒p3 -V2转化入E.COLIXL1-Blue中。结果 :应用电转化技术构建噬菌体抗体库库容为 :3.9×107单个scFv重组子 ,其转化效率为 :107～108 转化子/μg ,本实验最佳条件为场强 (7.5kv/cm) ,电容 (21.0μF) ,细菌浓度 (XL1 -Blue :4.0×1010/ml) ,外源DNA量 (p3 -V2:0.6μg)。结论 :经优化电转化条件 。Objective: To study the practical value of electrotransformation in constructing phage antibody repertoires has been Verified.Methods: Transform recombinant phagemid p3-V2 into E.COLI XL1-Blue strain using homebred gene pulser LN-101. Results: The library size: 3.9×107 individual scFv recombinants; the efficiency of transformation:107～108 transformants/ μg DNA added were obtained by setting the following parameters :field strength:7.5 kv/cm,capacitor:21.0 μF,recipient cell consistence :4.0×1010/ml,p3-V2 concentration:0.6 μg.Conclusion: After optimized electrotransformation factors, the results of this study meet the requirements of constructing a phage antibody repertoires.

关 键 词：电转化技术 噬菌体抗体库 库容 构建 

分 类 号：R392.11[医药卫生—免疫学] Q785[医药卫生—基础医学]