Molecular cloning and physical and translational mapping of the frog virus 3 genome

Abstract

A library of cloned fragments representing nearly the entire frog virus 3 (FV 3) genome (99.65%) has been constituted. Individual plasmid recombinants, labeled by nick-translation, were hybridized to Southern blots of genomic FV 3 DNA fragments obtained with XbaI, HindIII, SmaI, and SalI. From these results physical maps were generated and the distribution of restriction sites in the genome was established by double digestion of the fragments. A preliminary translational map was likewise developed. The viral messages were selected by hybridization to the recombinant DNAs immobilized on nitrocellulose filters and were translated in the reticulocyte cell-free system. About 30 polypeptides were detected among the translation products of RNA synthesized in the presence of cycloheximide. It appears that these genes are not clustered but in several cases more than one polypeptide is encoded by a given fragment. The 15 new polypeptide obtained by translation of late mRNAs derive from genes located on one-half of the genome.