伪狂犬病病毒Fa株gI和gp63基因缺失株的构建

Construction of pseudorabies virus Fa strain with deletions of glycoprotein gI and gP63 genes

用限制性核酸内切酶NcoI消化含有伪狂犬病病毒(PRV) Fa株Bam HI-7片段的质粒PBB7,以低融点琼脂糖回收目的片段, 经连接并转化E.coliDH5a, 获得缺失了gI和部分gp63基因的重组质粒PPB7-1。将PRVFa与PPB7-1DNA共同转染PK15单层细胞,待出现50% 以上细胞病变时收获病毒, 并以蚀斑法得到纯化重组病毒株, 命名为PFDI/D63。小鼠试验证实, 缺失株对小鼠具有一定的免疫原性。

Plasmid PPB7 is a recombinant of plasmid pBR322 and the BamHI 1 7 fragment of Pseudorabies Virus (PRV), from which the recombinant plasmid PPB7 1 deleted gI gene and part of gP63 gene were obtained by way of digestion with restriction endonucleotidase NcoI. By cotransfection of PRV Fa DNA and PPB7 1 DNA, a deletion mutant, designated PFDI/D63, was constructed with calcium phosphate transfection system. Inoculation of mice with 2.0×10 7PFU of the recombinant viruses revealed that mice were partly phylactic against PRV Fa containing 2 MLD.