Azab Walid, Kato Kentaro, Arii Jun, Tsujimura Koji, Yamane Daisuke, Tohya Yukinobu, Matsumura Tomio, Akashi Hiroomi
Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Arch Virol. 2009;154(5):833-42. doi: 10.1007/s00705-009-0382-0. Epub 2009 Apr 23.
Equine herpesvirus 4 (EHV-4) is a major cause of respiratory tract disease in horses worldwide. The generation of recombinant viruses, which would lead to understanding of viral gene functions, has been hindered by the absence of suitable cell lines and small-animal models of the infection. In the present study, the genome of EHV-4 strain TH20p was cloned as a stable and infectious BAC without any deletions of the viral genes. Mini F plasmid sequences flanked by loxP sites were inserted into the intergenic region between genes 58 and 59. Coinfection of the recombinant virus with a recombinant adenovirus expressing Cre recombinase resulted in the excision of the BAC sequences. Importantly, the resulting recombinant EHV-4 replicated comparably to the wild-type virus in fetal horse kidney cells. The recombinant EHV-4 will facilitate EHV-4 research and provide the opportunity to exploit the power of BAC technology for production of recombinant viral vaccines.
马疱疹病毒4型（EHV-4）是全球马匹呼吸道疾病的主要病因。由于缺乏合适的细胞系和感染的小动物模型，阻碍了重组病毒的产生，而重组病毒的产生有助于了解病毒基因功能。在本研究中，EHV-4毒株TH20p的基因组被克隆为稳定且具有感染性的细菌人工染色体（BAC），病毒基因无任何缺失。两侧带有loxP位点的微型F质粒序列被插入到基因58和59之间的基因间隔区。重组病毒与表达Cre重组酶的重组腺病毒共感染导致BAC序列被切除。重要的是，产生的重组EHV-4在胎马肾细胞中的复制与野生型病毒相当。重组EHV-4将促进EHV-4的研究，并为利用BAC技术生产重组病毒疫苗提供机会。
