摘要
为了建立蛋白质亲和配基的高效筛选方法,以淀粉酶为靶分子,利用交替洗脱法从七肽噬菌体展示库中筛选具有高亲和力的噬菌体配体.结果表明,交替洗脱法筛选出的特异性配体的回收率和ELISA信号值均优于酸洗脱法;采用交替洗脱法能够更加有效和迅速地筛选到淀粉酶的高亲和力配体.分析筛选得到的不同噬菌体克隆DNA插入片断的氨基酸序列,发现了可能与配体高亲和性有关的氨基酸残基.
To establish an high effective method for screening affinity ligands for protein separation, alpha-amylase was used as a target molecule for screening its affinity ligands from heptapeptide phage display library by the alternating elution method. Compared with the acid screening method, the modified strategy yielded higher peptide recovery and the selected phage clones showed higher signals of enzyme-linked immunosorbent assay(ELISA). The results indicate that specific ligands of high affinity for alpha-amylase can be efficiently obtained by the alternating elution method. The amino acid sequences of the affinity clones are determined by DNA sequencing, and the key amino acids binding to alpha-amylase are analyzed and discussed by the motif similarity between the affinity peptides.
出处
《天津大学学报（自然科学与工程技术版）》
EI
CAS
CSCD
北大核心
2005年第2期159-162,共4页
Journal of Tianjin University：Science and Technology
基金
国家自然科学基金资助项目(20476081).
关键词
噬菌体展示库
淀粉酶
配体
交替洗脱
phage display library
amylase
affinity ligand
alternating elution
