福建农业学报2024，Vol.39Issue(9)：1035-1043,9.DOI:10.19303/j.issn.1008-0384.2024.09.004
猪传染性胃肠炎病毒S、N基因DNA疫苗载体构建及其免疫原性
Vector Construction and Immunogenicity of S and N Gene DNA Vaccine for TGEV
摘要
Abstract
[Objective]DNA vaccine vector of S and N genes of porcine transmissible gastroenteritis virus(TGEV)was constructed with the vaccine immunogenicity determined to pave the way for studying,preventing,and controling TGE.[Method]A and D sites on S and N from a TGEV were amplified.The N gene alone as well as the A and D sites fusion were cloned into the vaccine vector pCDNA3.1-His-C.Bioinformatics software was used to predict and analyze the secondary structure,tertiary configuration,subcellular localization,and dominant B cell epitope of S(A-D)and N proteins.The recombinant vectors were transfected into PK-15 cells,and expression distribution of N and the A and D sites fusion detected by indirect immunofluorescence and confocal detection.Mice were immunized with the single or combined recombinant vaccine vector to detect the IgG antibody using indirect ELISA.[Result]The A and D sites of the S were 498 bp and 606 bp,respectively,and the N,1149 bp in length.The nucleic acid vaccine expression vectors p-S(A-D)-His and p-N-His for the A and D sites(fusion)and N were constructed.Bioinformatics software predicted that,when TGEV infected the host cells,N protein was mainly located in the nucleus and mitochondria and S(A-D)largely in the cytoplasm and mitochondria,while S(A-D)had 7 and N,8 dominant B cell epitopes.All p-S(A-D)-His and p-N-His were successfully expressed in PK-15 cells distributed in the nucleus and cytoplasm.The immunized mice showed an effect of immunity in the order of p-N-His＞p-S(A-D)-His+p-N-His＞p-S(A-D)-His.[Conclusion]The DNA vaccine vectors of S and N of TGEV were successfully constructed.Strong specific antibodies were generated in lab mice after the immunization.关键词
猪传染性胃肠炎病毒/S基因/N基因/载体构建/免疫原性Key words
transmissible gastroenteritis virus/S gene/N gene/vector construction/immunogenicity分类
农业科技引用本文复制引用
成伟伟,容维中,杨明,李元新,赵子惠,陈伯祥,王佳,周瑶..猪传染性胃肠炎病毒S、N基因DNA疫苗载体构建及其免疫原性[J].福建农业学报,2024,39(9):1035-1043,9.基金项目
甘肃省青年科技基金计划项目(21JR7RA719) （21JR7RA719）
甘肃省农业生物技术研究与应用开发项目(GNSW-2011-24) （GNSW-2011-24）
甘肃省重点研发计划项目(18YF1NA021-1) （18YF1NA021-1）
