天然鼠源噬菌体单链抗体库的构建

畜牧兽医学报 ›› 2005, Vol. 36 ›› Issue (1): 98-99.

天然鼠源噬菌体单链抗体库的构建

邵建军;苗向阳;朱瑞良;丁淑燕;陈永福

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

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2005-01-25 发布日期:2005-01-25

Construction of Naive Mouse Phagemid Single Chain Antibody Library

SHAO Jian-jun; MIAO Xiang-yang; ZHU Rui-liang; DING Shu-yan;CHEN Yong-fu

1. Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100094,China;2.College of Animal Science and Technology,Shandong Agricultural University,Taian 271018,China;3.The State Key Laboratories for AgroBiotechnology,China Agricultural University, Beijing 100094,China

Received:1900-01-01 Revised:1900-01-01 Online:2005-01-25 Published:2005-01-25

摘要/Abstract

Abstract: 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_Lwere 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.

Key words: phage antibody library, ScFv, amplification

邵建军;苗向阳;朱瑞良;丁淑燕;陈永福. 天然鼠源噬菌体单链抗体库的构建[J]. 畜牧兽医学报, 2005, 36(1): 98-99.

SHAO Jian-jun;MIAO Xiang-yang;ZHU Rui-liang;DING Shu-yan;CHEN Yong-fu. Construction of Naive Mouse Phagemid Single Chain Antibody Library[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2005, 36(1): 98-99.