VP22是一种保守的被膜蛋白,广泛存在于伪狂犬病病毒(PRV)和HSV-1等α-疱疹病毒中。HSV-1的VP22是病毒的重要毒力因子,它通过调节病毒蛋白和宿主蛋白的亚细胞定位,增强病毒的神经毒性;HSV-1 VP22还能够抑制AIM2炎症小体的激活,进而促进...VP22是一种保守的被膜蛋白,广泛存在于伪狂犬病病毒(PRV)和HSV-1等α-疱疹病毒中。HSV-1的VP22是病毒的重要毒力因子,它通过调节病毒蛋白和宿主蛋白的亚细胞定位,增强病毒的神经毒性;HSV-1 VP22还能够抑制AIM2炎症小体的激活,进而促进病毒的复制。然而,PRV VP22与HSV-1 VP22的同源性仅为26%,其在病毒感染和致病中的作用尚不明确。近期,mBio杂志发表了题为“Suppression of ZBP1-mediated NLRP3 inflammasome by the tegument protein VP22 facilitates pseudorabies virus infection”的研究论文,论文的通讯作者为南京农业大学动物医学院姜平教授和刘星教授,第一作者为博士后马梓承。本研究揭示了ZBP1介导的NLRP3炎症小体在PRV感染中的作用,并发现PRV VP22通过抑制该途径介导的炎症反应,促进病毒的复制和致病性。该研究成果为理解PRV复制和致病机制提供了新的视角。展开更多
To construct the bi-valent genetic engineering vaccine against pseudorabies virus（PRV）and porcine reproductive and respiratory syndrome virus（PRRSV）,the modified PRRSV ORF5 gene（ORF5M） and the VP22 gene of bovin...To construct the bi-valent genetic engineering vaccine against pseudorabies virus（PRV）and porcine reproductive and respiratory syndrome virus（PRRSV）,the modified PRRSV ORF5 gene（ORF5M） and the VP22 gene of bovine herpesvirus 1（BHV-1）,which encodes VP22 protein and has been demonstrated to exhibit the unusual protein transduction property,were inserted into a PRV universal transfer vector pIECMV by turns.A recombinant virus transfer vector pIECMV-VP22ORF5M possessing VP22-ORF5M fusion gene was generated.The recombinant virus transfer vector pIECMV-VP22ORF5M co-transfected the IBRS-2 cells with PRV TK-/gE-/LacZ+ genomic DNA digested by EcoRⅠusing liposome method.Based on homologous recombination,the recombinant virus was generated and then purified by the plaque assay and PCR amplification.After three rounds of plaque purification,the recombinant virus was further confirmed by PCR,Southern blot and Western blot.A recombinant PRV（rPRV）TK-/gE-/VP22GP5+ expressing VP22-GP5 fusion protein was constructed.The results of TCID50 tests showed that the insertion of the foreign genes had no influence on the propagation of rPRV in IBRS-2 or PK-15 cells.The construction of rPRV TK-/gE-/VP22GP5+ provides a basis for further study of bi-valent genetic engineering vaccines against PRRSV and PRV,and that this strategy may also be useful to develop more efficient genetic engineering vaccines against other pathogens.展开更多
文摘VP22是一种保守的被膜蛋白,广泛存在于伪狂犬病病毒(PRV)和HSV-1等α-疱疹病毒中。HSV-1的VP22是病毒的重要毒力因子,它通过调节病毒蛋白和宿主蛋白的亚细胞定位,增强病毒的神经毒性;HSV-1 VP22还能够抑制AIM2炎症小体的激活,进而促进病毒的复制。然而,PRV VP22与HSV-1 VP22的同源性仅为26%,其在病毒感染和致病中的作用尚不明确。近期,mBio杂志发表了题为“Suppression of ZBP1-mediated NLRP3 inflammasome by the tegument protein VP22 facilitates pseudorabies virus infection”的研究论文,论文的通讯作者为南京农业大学动物医学院姜平教授和刘星教授,第一作者为博士后马梓承。本研究揭示了ZBP1介导的NLRP3炎症小体在PRV感染中的作用,并发现PRV VP22通过抑制该途径介导的炎症反应,促进病毒的复制和致病性。该研究成果为理解PRV复制和致病机制提供了新的视角。
文摘To construct the bi-valent genetic engineering vaccine against pseudorabies virus（PRV）and porcine reproductive and respiratory syndrome virus（PRRSV）,the modified PRRSV ORF5 gene（ORF5M） and the VP22 gene of bovine herpesvirus 1（BHV-1）,which encodes VP22 protein and has been demonstrated to exhibit the unusual protein transduction property,were inserted into a PRV universal transfer vector pIECMV by turns.A recombinant virus transfer vector pIECMV-VP22ORF5M possessing VP22-ORF5M fusion gene was generated.The recombinant virus transfer vector pIECMV-VP22ORF5M co-transfected the IBRS-2 cells with PRV TK-/gE-/LacZ+ genomic DNA digested by EcoRⅠusing liposome method.Based on homologous recombination,the recombinant virus was generated and then purified by the plaque assay and PCR amplification.After three rounds of plaque purification,the recombinant virus was further confirmed by PCR,Southern blot and Western blot.A recombinant PRV（rPRV）TK-/gE-/VP22GP5+ expressing VP22-GP5 fusion protein was constructed.The results of TCID50 tests showed that the insertion of the foreign genes had no influence on the propagation of rPRV in IBRS-2 or PK-15 cells.The construction of rPRV TK-/gE-/VP22GP5+ provides a basis for further study of bi-valent genetic engineering vaccines against PRRSV and PRV,and that this strategy may also be useful to develop more efficient genetic engineering vaccines against other pathogens.
