Production of monoclonal antibodies against AFLD(Nor-1),an early key protein involved in aflatoxin biosynthesis  

作　　者：Ting Wang Peiwu Li Qi Zhang Wen Zhang Zhaowei Zhang Xueli Dong Tong Wang 

机构地区：[1]Key Laboratory of Biology and Genetic Improvement of Oil Crops,Ministry of Agriculture,Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences [2]Key Laboratory of Detection for Mycotoxins,Ministry of Agriculture [3]Laboratory of Risk Assessment for Oilseeds Products,Wuhan,Ministry of Agriculture [4]Quality Inspection and Test Center for Oilseeds Products,Ministry of Agriculture

出　　处：《Oil Crop Science》2018年第1期50-56,共7页中国油料作物学报(英文版）

基　　金：supported by National Natural Science Foundation of China (31471650, 31640062);National Key Research and Development Program of China (2016YFE0119900)

摘　　要：Aflatoxins are distributed worldwide throughout environment. As many as 30genes are involved in aflatoxin biosynthesis. aflD (nor-1) was predicted to encode a 31 kDa ketoreductase (AFLD), which catalyzed the earliest and first stable aflatoxin precursor in aflatoxin biosynthetic pathway. AFLD was proved to be necessary for conversion of norsolorinic acid (NA) to averatin (AVN) in aflatoxin biosynthesis. For this reason, aflD was cloned and specific monoclonal antibodies for AFLD were developed to better define potential pathways of AFLD involved in Aflatoxin B_1 (AFB_1) biosynthesis. Monoclonal antibodies 2A8-1E10 and 2F11-3D8 were successfully screened out by immunized mouse. Immunoblot analysis revealed that 2A8-1E10 had high sensitivity and specificity to identify native AFLD protein in A. flavus with detection limit of 3 ng/mL. These results showed that it was suitable for quantitative detection of AFLD in A. flavus isolate. Further investigation revealed that aflatoxin accumulations of various A. flavus were not dependent on AFLD biosynthesis.Overall, this is the first report for AFLD monoclonal antibody development and application in A. flavus quantitative detection.Aflatoxins are distributed worldwide throughout environment. As many as 30genes are involved in aflatoxin biosynthesis. aflD (nor-1) was predicted to encode a 31 kDa ketoreductase (AFLD), which catalyzed the earliest and first stable aflatoxin precursor in aflatoxin biosynthetic pathway. AFLD was proved to be necessary for conversion of norsolorinic acid (NA) to averatin (AVN) in aflatoxin biosynthesis. For this reason, aflD was cloned and specific monoclonal antibodies for AFLD were developed to better define potential pathways of AFLD involved in Aflatoxin B_1 (AFB_1) biosynthesis. Monoclonal antibodies 2A8-1E10 and 2F11-3D8 were successfully screened out by immunized mouse. Immunoblot analysis revealed that 2A8-1E10 had high sensitivity and specificity to identify native AFLD protein in A. flavus with detection limit of 3 ng/mL. These results showed that it was suitable for quantitative detection of AFLD in A. flavus isolate. Further investigation revealed that aflatoxin accumulations of various A. flavus were not dependent on AFLD biosynthesis.Overall, this is the first report for AFLD monoclonal antibody development and application in A. flavus quantitative detection.

关 键 词：AFLATOXIN BIOSYNTHESIS AFLD MONOCLONAL ANTIBODIES QUANTITATIVE detection 

分 类 号：S[农业科学]