Angeletti Peter C
Nebraska Center for Virology, School of Biological Sciences, University of Nebraska-Lincoln, USA.
Methods Mol Med. 2005;119:247-60. doi: 10.1385/1-59259-982-6:247.
Improvements in methodologies to recapitulate and study particular biological functions of the papillomavirus life cycle have led to great advances in our knowledge of these viruses. Described in this chapter are techniques that allow low-copy and high-copy replication of full-length human papillomavirus (HPV) genomes, as well as assembly of virus-like particles, in Saccharomyces cerevisiae (yeast). This system has several distinct advantages that make it an attractive complement to the well-established raft-culturing system. First, yeast are inexpensive, rapid, and simple to culture in the lab. Second, they provide an ever-widening array of genetic tools to analyze HPV functions--most recently notable, the yeast open reading frame (ORF)-deletion library. Third, yeast provide a potentially high-efficiency means to produce large quantities of infectious virus in a short time frame. Fourth, assembly of HPV virus in yeast allows encapsidation of mutant genomes, since previous studies have shown that no viral ORF is required for replication of full-length HPV in yeast.
用于概括和研究乳头瘤病毒生命周期特定生物学功能的方法学改进,使我们对这些病毒的认识取得了巨大进展。本章介绍的技术能够在酿酒酵母(酵母)中实现全长人乳头瘤病毒(HPV)基因组的低拷贝和高拷贝复制,以及病毒样颗粒的组装。该系统具有几个明显的优势,使其成为成熟的筏式培养系统的有吸引力的补充。首先,酵母在实验室中培养成本低、速度快且操作简单。其次,它们提供了越来越多的遗传工具来分析HPV功能——最近特别值得注意的是酵母开放阅读框(ORF)缺失文库。第三,酵母提供了一种潜在的高效方法,能够在短时间内大量生产感染性病毒。第四,HPV病毒在酵母中的组装允许突变基因组的衣壳化,因为先前的研究表明,酵母中全长HPV的复制不需要病毒ORF。