动物医学进展2024，Vol.45Issue(9)：1-6,6.
猪繁殖与呼吸综合征病毒重组N蛋白原核表达及间接ELISA方法的建立
Prokaryotic Expression of Recombinant PRRSV N Protein and Establishment of Indirect ELISA Method
摘要
Abstract
The study was aimed to prepare polyclonal antibody against PRRSV N protein and develop indi-rect ELISA to detect antibody against PRRSV N protein.The gene encoding PRRSV N protein was ampli-fied and cloned into pET-32a(+)prokaryotic expression vector,resulting in the pET-32a-N recombinant plasmid.The recombinant plasmid was deliveried into BL21(DE3)cells.The expression of N protein was in-duced by IPTG and then purified.The polyclonal antibodies were obtained by immunizing mice with recom-binant N protein.The indirect ELISA method for detecting antibodies to PRRSV N protein was developed and the specificity,reproducibility and accuracy of the method were also evaluated by using purified N pro-tein as antigen.The results showed that the expressed recombinant N protein was soluble and the poly-clonal antibody potency of 1∶2 048 000 was measured by indirect ELISA.Furthermore,both IFA and Western blot results showed that the prepared murine polyclonal antibody could specifically recognize PRRSV N protein.The indirect ELISA method based on the expressed N protein showed good specificity,the coefficient of variation of the reproducibility test was less than 10％,and the compliance rate with the re-sults of the same type of commercial kits reached 91.10％.The results of the study provide an experimental basis for the immunological detection of PRRSV N proteins and further studies of their structure and function.关键词
猪繁殖与呼吸综合征病毒/N蛋白/原核表达/多克隆抗体/间接ELISAKey words
Porcine reproductive and respiratory syndrome virus/N protein/prokaryotic expression/poly-clonal antibody/indirect ELISA分类
农业科技引用本文复制引用
覃建光,韦祖樟,姚静,刘文波,刘嘉琪,陈国昌,任同伟,陈樱,欧阳康,黄伟坚..猪繁殖与呼吸综合征病毒重组N蛋白原核表达及间接ELISA方法的建立[J].动物医学进展,2024,45(9):1-6,6.基金项目
国家自然科学基金项目(32172889,32370345) （32172889,32370345）
