【作者】 李祥敏； 钱平； 金梅林； 郭东春； 徐卓菲； 陈焕春；

【Author】 LI Xiang-min, QIAN Ping, JIN Mei-lin, GUO Dong-chun, SHU Zhuo-hui,CHEN Huan-chun(Laboratory of Animal Virology, College of Animal Husbandry and Veterinary Science, Hua Thong Agricultural University, Wu han 430070, China)

【机构】 华中农业大学畜牧兽医学院动物病毒室；

【摘要】 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(Foot-and-mouth disease virus,FMDV)P1基因插入到载体pPgG-uni中,得到伪狂犬病毒病毒(pseudorabies virus,PRV)转移载体pPgG-P1。将转移载体与EcoRI线性化后的PRV弱毒疫苗株TK/gG~-/lacZ~+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~-/gG~-/PgG-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伪狂犬病病毒弱毒疫苗株的基因组中。Western blot试验表明FMDV P1基因在重组伪狂犬病病毒中都得到表达。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。更多还原

【Abstract】 To construct the bivalent 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 co-transfected with the genome DNA of PRY 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 purified 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’s very useful to research the genetic engineering vaccine against PRV and FMDV in the future.更多还原