[Cloning and infectivity analysis of the cDNAs of tobacco mosaic virus (tomato strain) and its attenuated virus(N14) genomes].
An oligonucletide primer with SP6 transcription promoter and the other primer with a KpnI site at 5' terminuses were used to prepare the cDNAs of tobacco mosaic virus(TMV, Chinese isolate) and its attenuated virus(N14, nitrite-treated mutant, tomato strain) genomic RNAs by RT-PCR. The cDNAs were cut into two fragments by BamHI and first cloned separately then recombined into pUC18 by the restriction endonucleases of BamHI, SmaI, HincII, PstI, SphI. The sequences of 5' and 3' terminal nucleotides (< 50 bp) were consistent with the reported references. The infectious clones(pTMV-Cv/pN14) have been obtained and corroborated by in vitro transcription with SP6 RNA polymerase and tobacco infection. Necrotic local lesions were observed in Nicotiana samsun NN induced by the transcripts from both pTMV and pN14 within 3-5 days after inoculation, systemic symptoms showed in single-lesion infected Nicotiana tobacum cv, coincided with symptoms of tobaccoes infected by TMV and N14.
Yang
G
G
Institute of Microbiology, Chinese Academy of Sciences, Beijing.
Qiu
B S
BS
D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis* / Q000737:chemistry; D016679:Genome, Viral; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014027:Tobacco Mosaic Virus / Q000235:genetics*
D018076:DNA, Complementary