Virology, Nebraska Center for


Date of this Version



Methods Mol Med. 2005 ; 119: 247–260.


Copyright 2005 Angeletti.


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 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.