Position: Home > Articles > Screening and identification of Cry2Aa-binding peptides from a phage 12-mer peptide library
Jiangsu Journal of Agricultural Sciences
2015
(4)
929-934
Cry2Aa毒素结合十二肽的筛选与鉴定
作 者：
武爱华;张霄;刘媛;赵扬;赵岩岩
单 位：
江苏省农业科学院食品质量安全与检测研究所;南京农业大学植物保护学院江苏省农业科学院食品质量安全与检测研究所/江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地/农业部农产品量安全控制技术与标准重点实验室河南科技学院食品学院;江苏省农业科学院食品质量安全与检测研究所/农业部食品质量安全监控重点开放实验室南京农业大学植物保护学院;江苏省食品质量安全重点实验室和农业部食用农产品安全监控重点开放实验室;西北农林科技大学食品科学与工程学院
关键词：
Cry2Aa毒素;噬菌体展示肽库;ELISA
摘 要：
利用Cry2Aa毒素蛋白对噬菌体展示随机十二肽库进行4轮筛选,并从第4轮筛选产物中随机挑选20个单克隆,进行单克隆ELISA、PCR扩增、DNA电泳及测序,鉴定这些克隆与Cry2Aa毒素的结合活性。结果显示,这20个克隆均能与Cry2Aa毒素发生特异性结合,并推导出8条不同的序列。挑取阳性值最高的克隆GT(氨基酸序列为GTPWHHHRHLIV)建立了基于十二肽的Cry2Aa毒蛋白的间接竞争ELISA检测方法。该方法的抑制中浓度(IC50)为1.139 0μg/ml,最低检测限IC10为0.021 1μg/ml,线性检测范围为0.047 8~22.691 0μg/ml(Y=23.09 lgx+48.692,R2=0.996 3)。噬菌体展示肽库筛选方法为快速、准确地检测Cry2 Aa毒蛋白提供了新的途径。
译 名：
Screening and identification of Cry2Aa-binding peptides from a phage 12-mer peptide library
作 者：
WU Ai-hua;ZHANG Xiao;LIU Yuan;ZHAO Yang;ZHAO Yan-yan;ZHANG Cun-zheng;XIE Ya-jing;LIU Xian-jin;Institute of Food Quality Safety and Detection,Jiangsu Academy of Agricultural Sciences/Key Laboratory of Control Technology and Standard for Agroproduct Safety and Quality,Ministry of Agriculture;College of Plant Protection,Nanjing Agricultural University;
关键词：
Cry2Aa toxin;;phage-display peptide library;;ELISA
摘 要：
A phage displayed random 12-mer peptide library( Ph. D.-12 TM Phage Display Peptide Library) is screened with Cry2Aa toxin. After 4 rounds of panning,20 single colonies were selected randomly,and the positive clones are identified by monoclonal phage competitive enzyme-linked immunosorbent assay( ELISA),and PCR,DNA electrophoresis and sequencing. Totally 8 peptides,which could recognize Cry2Aa specifically have been confirmed. The peptide( amino acid sequence: GTPWHHHRHLIV) holding better binding ability was employed to develop an indirect competitive ELISA for the Cry2Aa detection. The peptide-based indirect competitive ELISA showed that the IC50 was 1. 139 0 μg / ml,the minimum detection limit was 0. 021 1 μg / ml,and the linear detection ranged from 0. 047 8 to 22. 691 0 μg / ml. With this new detection method,Cry2Aa can be detected rapidly and accurately.
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