Infectious positive- and negative-strand transcript RNAs from bacteriophage Q beta cDNA clones.
Plasmids containing full-length cDNA copies of the Q beta RNA phage genome and flanking T7 promoters were constructed. Positive-strand Q beta RNA, generated by in vitro transcription of these plasmids with T7 RNA polymerase, was infectious to Escherichia coli spheroplasts. The Q beta replicase gene from the cloned DNA was subcloned and expressed in E. coli cells by means of a thermoinducible plasmid. Full-length, negative-strand Q beta transcripts were infectious when transfected into spheroplasts containing the induced replicase gene.
Shaklee
P N
PN
Biochemistry Department, University of Wisconsin, Madison 53706.
Miglietta
J J
JJ
Palmenberg
A C
AC
Kaesberg
P
P
D003001:Cloning, Molecular; D003090:Coliphages / Q000235:genetics*; D004247:DNA / Q000235:genetics; D004926:Escherichia coli; D005814:Genes, Viral; D012367:RNA, Viral / Q000235:genetics*; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics; D013104:Spheroplasts; D014158:Transcription, Genetic; D014162:Transfection
D012367:RNA, Viral; D004247:DNA; D012324:RNA-Dependent RNA Polymerase