牛Ⅰ型疱疹病毒感染性细菌人工染色体的构建和<italic>gN</italic>基因缺失病毒的细胞繁殖特性
摘要
The recombinant bovine herpesvirus type 1 (rBHV1-HA) was constructed by inserting pHA2 plasmid into the viral genome between UL15 and UL18 cassettes. BHV1 infectious bacterial artificial chromosome had been confirmed after rBHV1-HA circular genome was extracted and transformed into <italic>E. coli</italic> strain DH10B. The resulting Bac clone, pBHV-1, was transfected into bovine kidney cells (MDBK) and the virus was rescued. The rescued virus, BHV1-res, have almost the same titres as the wildtype in replication <italic>in vitro</italic>. The transmembrane domain in <italic>gN</italic> open reading fram was deleted from pBHV-1 by Red E/T mutagenesis in <italic>E.coli</italic>. BHV-1 with partial deletion of <italic>gN</italic>, rBHV1-?gN, was reconstituted from the mutated Bac. rBHV1-?gN had lower titres by from 9% to 20% than rBHV-res in growth property in MDBK. This construction of infectious BHV1 clone should contribute greatly to the recombinant BHV1 vaccine with genes deletion and bovine universal viral vector.