Functional genomics on potato virus A: virus genome-wide map of sites essential for virus propagation.

ABSTRACT: Transposition-based in vitro insertional mutagenesis strategies provide promising new approaches for functional characterization of any cloned gene or genome region. We have extended the methodology and scope of such analysis to a complete viral genome. To map genome regions both essential and nonessential for Potato virus A propagation, we generated a genomic 15-bp insertion mutant library utilizing the efficient in vitro DNA transposition reaction of phage Mu. We then determined the proficiency of 1125 mutants to propagate in tobacco protoplasts by using a genetic footprinting strategy that simultaneously mapped the genomic insertion sites. Over 300 sites critical for virus propagation were identified, and many of them were located in positions previously not assigned to any viral functi

SUBMITTER: Kekarainen T 

PROVIDER: S-EPMC187510 | biostudies-literature | 2002 Apr

REPOSITORIES: biostudies-literature