动物医学进展2025，Vol.46Issue(11)：50-55,6.
赤羽病毒N基因的序列分析及原核表达
Sequence Analysis and Prokaryotic Expression of the N Gene of Akabane Virus
摘要
Abstract
To investigate the genetic variation of the N gene of the Akabane virus(AKAV)and to prepare detection antigens using prokaryotic expression technology,primers were designed based on the N genome sequence of AKAV available in GenBank.After reverse transcription of AKAV RNA,the target gene was amplified using PCR.Following the sequencing of the target fragment,comprehensive analyses were con-ducted regarding the physicochemical properties of the protein,gene sequence,and prokaryotic expression.The results indicated that the 717 bp N gene of AKAV was successfully amplified.The encoded protein was stable,lacking transmembrane region and signal peptides,and exhibited hydrophilic and alkaline prop-erties.Nucleotide sequence analysis revealed that the homology of the N gene sequence of this strain,com-pared to those published in NCBI,ranged from 82.7％to 97.9％.The prokaryotic expression plasmid pET28a-N was constructed and subsequently transformed into the BL21 strain,resulting in the successful expression of the 27 ku recombinant N protein.Western blot analysis confirmed that this protein could bind to positive serum.This experiment laid the foundation for the further development of AKAV vaccines and ELISA diagnostic kits.关键词
赤羽病毒/序列分析/核衣壳基因/原核表达Key words
Akabane virus/N gene/Sequence analysis/Prokaryotic expression引用本文复制引用
马勇杰,王凯茸,刘建行,张淑霞,张琪,许信刚..赤羽病毒N基因的序列分析及原核表达[J].动物医学进展,2025,46(11):50-55,6.基金项目
国家重点研发计划(2021YFD1600704) （2021YFD1600704）
省级农业专项资金项目(XNDY2210) （XNDY2210）
