赭曲霉毒素A噬菌体单链抗体库的构建与鉴定
首发时间:2015-04-27
摘要:从赭曲霉毒素A(OTA)免疫小鼠的脾细胞中扩增出VH和VL 基因片段,再经重叠延伸PCR( SOE-PCR) 拼接扩增得到单链抗体( ScFv) 基因,然后克隆到pCANTAB5E噬菌粒载体上,转化感受态大肠杆菌TG1,并经辅助噬菌体M13K07超感染,构建OTA毒素噬菌体单链抗体库,ScFv基因插入率为100%。免疫亲和筛选和富集后,经ELISA法最终筛选得到5株可分泌阳性噬菌体抗体的细菌 结论: 赭曲霉毒素A 噬菌体单链抗体库构建成功。
关键词: 赭曲霉毒素A; 单链抗体; 噬菌体抗体库
For information in English, please click here
Construction and identification of phage ScFv library against Ochratoxin A
Abstract:The VH and VL gene fragments were first amplified from spleen cells of mice specific immune to Ochratoxin A(OTA) integrated as single-chain antibody(ScFv) gene using gene splicing by overlap extension PCR( SOE-PCR).The ScFv was cloned into a phagemid vector pCANTAB5E,and then was transformed into Escherichia coli TG1 to establish a phage ScFv library with the helper phage M13K07.The results indicated that the insert rate of SvFv gene was 100%. After biopanning enrichment and screening,five strains that could secrete the FB1 specific phage single-chain antibody were selected successfully.Conclusion:the phage ScFv library against FB1 was constructed successf
Keywords: Ochratoxin A single-chain antibody( ScFv) phage ScFv library
论文图表:
引用
No.4639842105175214****
同行评议
共计0人参与
勘误表
赭曲霉毒素A噬菌体单链抗体库的构建与鉴定
评论
全部评论0/1000