Genomic nucleotide sequence of a foot-and-mouth disease virus clone and its persistent derivatives

Distance effects during polyprotein processing in the complementation between defective FMDV RNAs

“…The origin of baby hamster kidney 21 (BHK-21) cells and procedures for cell growth in Dulbecco's modification of Eagle's medium (DMEM), and for FMDV plaque assays in semisolid agar medium have been described previously (Domingo et al, 1980;Sobrino et al, 1983). FMDV C-S8c1 (expressed from plasmid pMT28; Toja et al, 1999) has the genomic sequence of a plaque-purified virus of the European serotype C, natural isolate C 1 Santa-Pau Spain 70 (Sobrino et al, 1983). FMDV C-S8p260 is a viral population obtained after 260 serial cytolytic passages of C-S8c1 at high m.o.i.…”

“…Expression of FMDV L. Plasmid pTM1-L was obtained with primers 5¢NcoI-L and 3¢BamHI-L, which were designed for PCR amplification of the Lab-coding region from pMT28 (Toja et al, 1999). The resulting membrane for protein visualization with the relevant antibodies.…”

Persistence of foot-and-mouth disease virus in cell culture revisited: implications for contingency in evolution

“…However, the consensus sequence of genomic residues 1897-3030 (residues encoding VP2 and part of VP3) and 6610-8019 (residues encoding 3D) could be determined, and they were compared with the consensus nucleotide sequence of VR100, the persistent virus shed by carrier cells at R cell passage 100 (Díez et al, 1990a;Toja et al, 1999). In the VP2-and VP3-coding regions examined, three mutations at genomic positions 2433, 2477 and 2517 that were not detected at passage 45 were dominant in the consensus sequence of R-D100 (Table 5).…”

“…Interestingly, none of the mutations present in the 3D-coding region at passage 45 were maintained as dominant at passage 100 and, furthermore, four additional mutations, three of them leading to amino acid substitutions in either R-B and R-D, were present in the consensus sequences at passage 100 (compare Tables 4 and 5). The comparison with the corresponding regions of FMDV VR100 (Díez et al, 1990a;Toja et al, 1999) indicates that two mutations (D3009A and N3013H) were common to the three lineages (R, R-B and R-D cells). All the phenotypic traits analysed and part of the genetic changes undergone by persistent FMDV point to a remarkable reproducibility of the coevolutionary events during FMDV persistence in BHK-21 cells, despite an inherent genetic instability of both the cells and the resident virus.…”

“…The main feature of FMDV persistence in cell culture was a rapid coevolution of the cells and the resident virus, reflected in a gradual increase of resistance of the cells to infection by the parental FMDV, and a gradual increase of FMDV virulence towards the parental BHK-21 cells (de la Torre et al, 1988Torre et al, , 1989aMartín Hernández et al, 1994;Sáiz & Domingo, 1996;Toja et al, 1999). These events conformed to the accepted definition of coevolution, meaning an interdependence of the evolution of two interacting biological entities (Futuyma & Slatkin, 1983;Woolhouse et al, 2002).…”