南方农业学报2017，Vol.48Issue(11)：2064-2070,7.DOI:10.3969/j.issn.2095-1191.2017.11.23
抗黄曲霉毒素B1单链抗体基因克隆及其结构分析
Anti-aflatoxin B1 single chain antibody:Gene cloning and structure analyzing
摘要
Abstract
[Objective]In this work,the anti-aflatoxin B1(AFB1)single chain antibody(scFv)gene was constructed, and its encoded protein sequence and structure was analyzed and predicted,which provided reference for scFv molecular modification and application in immunological detection.[Method]In this study,anti-AFB1 hybridoma cell strains were used as materials,the variable region of heavy chain(VH)and light chain(VL)were amplified by PCR,then anti-AFB1 gene scFv was built with(Gly4Ser)3 as flexible joint(Linker). The amino acid sequence,its physicochemical properties and protein structure were analyzed by online bioinformatics analysis software.[Result]The results indicated that the full anti-AFB1 gene scFv was 744 bp in length,encoding 248 amino acids. The molecular weight was 26312.05 Da,and theoretical isoelectric point(pI)5.70. In secondary structure,there were 35α-helixes(14.11%),28β-folds(11.29%),85 extended strands(34.28%)and 100 random coils(40.32%). In tertiary structure,connective peptides pulled together VH and VL to form a typical abtibidu structure—groove structure,which was the antigen binding region for scFv.[Conclu-sion]The constructed anti-AFB1 scFv VH contain 117 amino acids and VL contained 116 amino acids with a typical antibody structure—groove structure,which can combine with antigen specificity.关键词
黄曲霉毒素B1(AFB1)/单链抗体(scFv)/重链可变区(VH)/轻链可变区(VL)/沟槽结构Key words
aflatoxin B1(AFB1)/single chain antibody(scFv)/variable region of heavy chain(VH)/variable re-gion of light chain(VL)/groove structure分类
农业科技引用本文复制引用
马兰,刘爱平,王佳,王小红..抗黄曲霉毒素B1单链抗体基因克隆及其结构分析[J].南方农业学报,2017,48(11):2064-2070,7.基金项目
国家自然科学基金项目(31601539) （31601539）
湖北省自然科学基金项目(2016CFB143) （2016CFB143）
华中农业大学自主科技创新基金项目(2662015QD027) （2662015QD027）
