天然鼠源噬菌体单链抗体库的构建  

Construction of Naive Mouse Phagemid Single Chain Antibody Library

作　　者：邵建军[1] 苗向阳[2] 朱瑞良[1] 丁淑燕[1] 陈永福[3] 

机构地区：[1]山东农业大学动物科技学院,泰安271018 [2]中国农业科学院畜牧研究所,北京100094 [3]中国农业大学农业生物技术重点实验室,北京100094

出　　处：《畜牧兽医学报》2005年第1期98-99,共2页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(30070543);北京市自然科学基金(5032012)资助

摘　　要：A murine phage antibody library was constructed.First,the total RNA was extracted from fresh spleens of nonimmunized mice,then the cDNA library was achieved via reverse transcription PCR.Gene fragments encoding V_H and V_L were amplified and assembled into a single gene using a DNA linker encoding a polypeptide of 15 amino acid residues(Gly4Ser)3 through PCR.And finally the recombinant DNA fragments were cloned into the phagemid pCANTAB5E vector and introduced into E.coli TG1.The phagemid particles displaying functional ScFv were rescued by reinfection of helper phage M13K07,thus a murine antibody library was obtained.A murine phage antibody library was constructed.First,the total RNA was extracted from fresh spleens of nonimmunized mice,then the cDNA library was achieved via reverse transcription PCR.Gene fragments encoding V_H and V_L were amplified and assembled into a single gene using a DNA linker encoding a polypeptide of 15 amino acid residues(Gly4Ser)3 through PCR.And finally the recombinant DNA fragments were cloned into the phagemid pCANTAB5E vector and introduced into E.coli TG1.The phagemid particles displaying functional ScFv were rescued by reinfection of helper phage M13K07,thus a murine antibody library was obtained.

关 键 词：单链抗体 丝状噬菌体 天然 动物体内 噬菌体抗体库 重链 噬菌体展示技术 DNA片段 体外表达 外源基因 

分 类 号：S852[农业科学—基础兽医学] R392[农业科学—兽医学]