伪狂犬病病毒Ea株TK^-/gE^-/gp63^-突变株的构建及其生物学特性研究  

Construction of Pseudorabies Virus Ea TK^-/gE^-/gp63^- MutantStrain and the Study on Its Biological Property

作　　者：刘正飞[1] 陈焕春[1] 何启盖[1] 周复春[1] 方六荣[1] 

机构地区：[1]农业部农业微生物重点实验室,华中农业大学,武汉430070

出　　处：《微生物学报》2002年第3期370-374,共5页Acta Microbiologica Sinica

基　　金：高等学校博士学科点专项科研基金资助项目;国家自然科学基金资助项目 ( 39970 5 5 9)~~

摘　　要：Using pseudorabies virus Ea strain as material,we inserted LacZ gene expression cassette into gE gene.After blue plaque and plaque purification,a recombinant virus PRVEa TK+-/gE+-/LacZ++ generated.Utilizing EcoR I site in LacZ gene, digested PRVEa TK+-/gE+-/LacZ++ genome DNA was cotransfected into PK-15 cells with plasmid pFBBS,then PRVEa TK+-/gE+-/gp63+- generated after plaque purification.Four pairs of primers amplification demonstrated the virus was pure TK+-/gE+-/gp63+- mutant virus.PCR product sequence indicates there were 205bp deletion in TK gene;1247bp deletion in gE,gp63 and intergenic region of PRVEa TK+-/gE+-/gp63+- mutant virus genome DNA.Inoculation to Balb/C mice with PRVEa TK+-/gE+-/gp63+- indicates the virulence is reduced greatly.Using pseudorabies virus Ea strain as material,we inserted LacZ gene expression cassette into gE gene.After blue plaque and plaque purification,a recombinant virus PRVEa TK+-/gE+-/LacZ++ generated.Utilizing EcoR I site in LacZ gene, digested PRVEa TK+-/gE+-/LacZ++ genome DNA was cotransfected into PK-15 cells with plasmid pFBBS,then PRVEa TK+-/gE+-/gp63+- generated after plaque purification.Four pairs of primers amplification demonstrated the virus was pure TK+-/gE+-/gp63+- mutant virus.PCR product sequence indicates there were 205bp deletion in TK gene;1247bp deletion in gE,gp63 and intergenic region of PRVEa TK+-/gE+-/gp63+- mutant virus genome DNA.Inoculation to Balb/C mice with PRVEa TK+-/gE+-/gp63+- indicates the virulence is reduced greatly.

关 键 词：伪狂犬病病毒EA株 TK^-/gE^-/gp63^-突变株 生物学特性 

分 类 号：S852.65[农业科学—基础兽医学]