Full-length biologically active cDNAs of brome mosaic virus genomic RNAs 1, 2 and 3 were constructed by joining cDNA fragments. The cDNAs were constructed so that, at the 5′ ends, unique SnaBI sites were present at the site of initiation of transcription. The cDNAs were inserted between a modified cauliflower mosaic virus (CaMV) 35S RNA promoter and terminator regions derived from CaMV DNA, and cloned into pUC18. When a mixture of the plasmid DNAs was inoculated onto Chenopodium hybridum leaves, local lesions appeared 5 to 6 days later. However, no symptoms appeared in similarly inoculated barley plants. Plasmid cDNAs with extra sequences at the 5′ end were infectious but RNAs transcribed from cDNAs with similar sequences were not.
AhlquistP.,
DasguptaR.,
KaesbergP.1984a; Nucleotide sequence of the brome mosaic virus genome and its implications for viral replication. Journal of Molecular Biology 172:369–383
AhlquistP.,
FrenchR.,
JandaM.,
Loesch-FriesL. S.1984b; Multicomponent RNA plant virus infection derived from cloned viral cDNA. Proceedings of the National Academy of Sciences, U,. S,. A 81:7066–7070
JandaM.,
FrenchR.,
AhlquistP.1987; High efficiency T7 polymerase synthesis of infectious RNA from cloned brome mosaic virus cDNA and effects of 5′ extensions on transcript infectivity. Virology 158:259–262
LaneL. C.1981; Bromoviruses. In Handbook of Plant Virus Infections: Comparative Diagnosis pp. 333–376 Edited by
KurstakE.
Amsterdam: Elsevier/North-Holland;
TaniguchiT.,
PalmieriM.,
WeissmannC.1978; Qβ DNA-containing hybrid plasmids giving rise to Qβ phage formation in the bacterial host. Nature, London 274:223–228
Van EmmeloJ. V.,
AmelootP.,
FiersW.1987; Expression in plant of the cloned satellite tobacco necrosis virus genome and of derived insertion mutants. Virology 157:480–487