Herpesvirus of turkey reconstituted from bacterial artificial chromosome clones induces protection against Marek's disease
Herpesvirus of turkey reconstituted from bacterial artificial chromosome clones induces protection against Marek's disease
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DOI：
10.1099/vir.0.81498-0
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发表时间：
2006-04-01
影响因子：
3.8
通讯作者：
Nair, VK
中科院分区：
文献类型：
--
作者：
Baigent, SJ;Petherbridge, LJ;Nair, VK
火鸡疱疹病毒（HVT）是一种α疱疹病毒，由于其与马立克氏病病毒（MDV）的抗原关系，被广泛用作马立克氏病的活疫苗。尽管有相似的基因组结构，HVT有几个独特的基因，其功能尚未完全了解。作为对HVT基因的功能进行详细分析的第一步，构建了HVT的全长感染性细菌人工染色体（BAC）克隆。来自两个独立BAC克隆的DNA在转染到鸡胚成纤维细胞中后，产生与野生型病毒产生的噬斑相似的噬斑。来自BAC克隆的病毒在体外传代期间是稳定的，但与野生型病毒相比在体外生长动力学上显示出差异。使用一步诱变，从HVT基因组中删除必需糖蛋白B基因的方案，随后构建回复突变体病毒，HVT的BAC克隆显示出适合于标准诱变技术。尽管在体外生长方面存在差异，但来自两个克隆的病毒诱导了100%的针对强毒MDV株RB-1B感染的保护，表明BAC-克隆化的病毒可用作疫苗，其效力与亲本病毒的效力相似。HVT BAC的构建是通过利用BAC技术的力量来理解HVT基因功能的重要一步。此外，BAC克隆的可用性使得能够使用HVT作为表达外源基因的载体。
Herpesvirus of turkey (HVT) is an alphaherpesvirus that is widely used as a live vaccine against Marek's disease because of its antigenic relationship with Marek's disease virus (MDV). In spite of a similar genome structure, HVT has several unique genes, the functions of which are not completely understood. As a first step in carrying out detailed analysis of the functions of the HVT genes, a full-length infectious bacterial artificial chromosome (BAC) clone of HVT was constructed. DNA from two independent BAC clones, upon transfection into chicken embryo fibroblasts, produced plaques similar to those produced by the wild-type virus. Viruses derived from the BAC clones were stable during in vitro passage, but showed differences in in vitro growth kinetics compared with the wild-type virus. Using a one-step mutagenesis, protocol to delete the essential glycoprotein B gene from the HVT genome, followed by construction of the revertant virus, BAC clones of HVT were shown to be amenable to standard mutagenesis techniques. In spite of the difference in in vitro growth, viruses from both clones induced 100 % protection against infection by the virulent MDV strain RB-1B, indicating that the BAC-clerived viruses could be used as vaccines with efficacies similar to that of the parental virus. The construction of HVT BAC is a major step in understanding the functions of HVT genes by exploiting the power of BAC technology. Furthermore, the availability of the BAC clones enables use of HVT as a vector for expressing foreign genes.