表达口蹄疫病毒P1基因的重组伪狂犬病病毒TK/gG-/PgG-P1的构建

为了构建口蹄疫与伪狂犬病二价基因工程疫苗株 ,将口蹄疫病毒 (FMDV) P1基因插入到伪狂犬病病毒 (PRV)通用载体 p Pg G- uni中 ,得到 PRV转移载体 p Pg G- P1。将转移载体与 Eco R 线性化后的 PRV弱毒疫苗株 TK- /g G- / lac Z 基因组 DNA共转染 PK- 15细胞 ,转染产物经多次空斑纯化和 PCR鉴定 ,获得了纯化的重组 PRV TK- /g G- / Pg G- P1,重组病毒基因组 DNA经酶切鉴定进一步表明 ,FMDV的 P1基因已成功地整合到 PRV弱毒疫苗株的基因组中。Western blot试验表明 ,FMDV P1基因在重组 PRV中得到表达。该研究为进一步研制口蹄疫与伪狂犬病二价基因工程疫苗奠定了坚实的基础

Construction of Recombinant Pseudorabies Virus (TK-/gG-/PgG-P1 ) Which Expressed P1 Gene of Foot-and Mouth Disease Virus

To construct the bi-valent genetic engineering vaccine against pseudorabies and foot-and mouth disease,P1 gene of foot-and mouth disease virus was inserted into a PRV universal vector-PgG-uni,and transfer vector PgG-P1 was constructed.The linear transfer vector was cotransfected with the genome DNA of PRV attenuated live vaccine strain(TK~-/gG~-/lacZ~ ) on PK-15 cells.The recombinant pseudorabies virus,designated TK~-/gG~-/PgG-P1,expressed FMDV P1 gene was obtained by three rounds of phage purification and PCR identification.FMDV P1 gene had been successfully inserted into the genome DNA of pseudorabies virus attenuated vaccine strain-TK~-/gG~-/lacZ~ by restriction enzyme analysis of DNA of the recombinant virus TK~-/gG~-/PgG-P1.Furthermore,the expression of FMDV P1 gene in recombinant virus was identified by Western blot.It would be very useful for researching the genetic engineering vaccine against PRV and FMDV in the future.