Synthesis of infectious transcripts of blueberry scorch carlavirus in vitro.
Blueberry scorch carlavirus (BBScV) is a filamentous virus with a polyadenylated, positive-sense RNA genome. A near full-length cDNA clone of BBScV was constructed by assembly of clones from a cDNA library. To generate a full-length cDNA clone, the 5' terminus was mutagenized by PCR to introduce nucleotides present in the wild-type virus and not in the near full-length clone, and then fused directly to the T7 bacteriophage RNA polymerase promoter at the 5' terminus. Capped and uncapped BBScV transcripts were synthesized in vitro from the full-length cDNA clone. Capped transcripts were infectious, producing systemic symptoms identical to those caused by the wild-type virus following inoculation onto Chenopodium quinoa leaves. Uncapped transcripts were substantially less infectious than capped transcripts. This represents the first report of infectious transcripts for a member of the carlavirus group.
Lawrence
D M
DM
Department of Plant Pathology, Rutgers University, Cook College, New Brunswick, New Jersey 08903.
Hillman
B I
BI
D001483:Base Sequence; D017788:Carlavirus / Q000378:metabolism*; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D005638:Fruit / Q000382:microbiology; D015870:Gene Expression; D016679:Genome, Viral; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011401:Promoter Regions, Genetic; D012367:RNA, Viral / Q000096:biosynthesis*; D014158:Transcription, Genetic
D017931:DNA Primers; D018076:DNA, Complementary; D012367:RNA, Viral; D012321:DNA-Directed RNA Polymerases