Transcription and translation maps of African swine fever virus

Abstract

A transcription map of African swine fever (ASF) virus DNA was obtained by hybridization of 32P-labeled early and late RNAs synthesized in Vero cells infected with ASF virus to dot-blots containing cloned restriction fragments spanning the viral genome. Early RNAs synthesized in infected cells in the presence of protein or DNA synthesis inhibitors hybridized preferentially to four regions in the genome, with coordinates E1 (0-51.9 kbp), E3 (63.7-75.2 kbp), E5 (100.1-111.6 kbp), and E7 (150-170 kbp). Late RNA present in infected cells after DNA replication hybridized with essentially all the genome. The RNA synthesized in vitro by the RNA polymerase associated with ASF virions hybridized to the same DNA regions than early RNA. After hybridization selection with DNA restriction fragments and translation in reticulocyte lysates the RNA synthesized in vitro produced the same proteins as early RNA. These results suggest that early RNA is synthesized in the infected cells by the virion-associated RNA polymerase. Maps of early and late proteins of ASF virus were constructed by cell-free translation of early or late RNAs selected by hybridization to cloned restriction fragments of virus DNA. About 100 early and 100 late polypeptide bands were mapped on the ASF virus genome.