Prokaryotic Expression of IBV N Protein and Development of Indirect IBV N Protein-mediated ELISA  

作　　者：Li Wei-qun Wang Xin Zhong Ming Sun Xiao-qi Zhao Lei Huang Xiao-dan Zhang Rui-li Li Guang-xing 无 

机构地区：[1]Key Laboratory for Laboratory Animals and Comparative Medicine of Heilongjiang Province,College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China [2]College of Pharmaceutical Engineering,Jilin Agriculture Science and Technology College,Jilin 132101,China

出　　处：《Journal of Northeast Agricultural University(English Edition)》2020年第1期69-79,共11页东北农业大学学报（英文版）

基　　金：Supported by the National Natural Science Foundation of China(31172295,31272569)。

摘　　要：Avian infectious bronchitis(IB)is an acute and highly contagious disease caused by infectious bronchitis virus(IBV).In the study,according to IBV gene sequences published in Gen Bank,specific primers were designed to clone N gene by RT-PCR,and this gene was inserted into p ET-30a(+)vector resulting in a prokaryotic expression plasma p ET-30a-N.The results of SDS-PAGE and Western Blot analysis showed that the recombinant protein was expressed successfully and had good reactivity with IBV positive serum.Using purified recombinant N protein as a coating antigen,the indirect ELISA protocol was established and optimized,in which N protein was 2.5μg·m L^-1 of concentration,sample serum of 1:40 dilution.For clinical specimen,the IBV antibodies could be detected by this method efficiently and got nearly the same results as those of IBV-mediated ELISA.It would provide a good tool for rapid diagnosis and epidemiological study of avian infectious bronchitis.

关 键 词：INFECTIOUS BRONCHITIS virus N protein PROKARYOTIC expression indirect ELISA 

分 类 号：S852.65[农业科学—基础兽医学]