食品科学2017，Vol.38Issue(10)：1-5,5.DOI:10.7506/spkx1002-6630-201710001
抗脱氧雪腐镰刀菌烯醇纳米抗体随机突变与筛选
Random Mutation and Mutant Screening of Anti-Deoxynivalenol Nanobody
摘要
Abstract
In this work,an anti-deoxynivalenol nanobody isolated from a naive phage display library was adopted to investigate the relationship between bioactivity and the composition of amino acid residues using error prone polymerase chain reaction (epPCR) and phage display.The epPCR was firstly optimized to produce a phage display library with balanced distribution of mutation.After six rounds of panning of the mutant library,30 out of 96 picked clones exhibited 8 different mutations,whose signal was 2.1 folds higher than that of the wild-type clone.Multi-sequence alignment analysis showed that mutant sites were prone to be found in the junction region between complementarity determining regions (CDRs) and framework regions (FRs).Three-dimensional structure modeling results indicated that some mutants were capable of forming an intramolecular disulphide bond,which benefits to stabilize the molecular structure.These data can provide a clue for further mutation and modification of anti-deoxynivalenol nanobody and also provide a guideline for the design of nanobody recognizing other low molecular weight compounds.关键词
纳米抗体/脱氧雪腐镰刀菌烯醇/易错聚合酶链式反应/噬菌体展示Key words
nanobody/deoxynivalenol/error prone PCR/phage display分类
轻工纺织引用本文复制引用
吴红静,曹冬梅,许杨,付金衡,涂追..抗脱氧雪腐镰刀菌烯醇纳米抗体随机突变与筛选[J].食品科学,2017,38(10):1-5,5.基金项目
国家自然科学基金青年科学基金项目(31301479) （31301479）
江西省教育厅科学技术研究项目(GJJ151502) （GJJ151502）
南昌大学科学技术学院自然科学基金项目(2013-ZR-05) （2013-ZR-05）
