伏马菌素B1噬菌体抗体库的构建与筛选
首发时间:2015-04-27
摘要:从伏马菌素B1( FB1) 杂交瘤细胞株F3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FB1 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: 伏马菌素B1噬菌体单链抗体库构建成功。
关键词: 伏马菌素B1; 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 fumonisin B1
Abstract:The VH and VL gene fragments were first amplified from hybridoma cell line( F3) specific to fumonisinB1(FB1) 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 successfully.
Keywords: fumonisin B1 single-chain antibody( ScFv) phage ScFv library
论文图表:
引用
No.4639839105175214****
同行评议
共计0人参与
勘误表
伏马菌素B1噬菌体抗体库的构建与筛选
评论
全部评论0/1000