The open reading frame of bamboo mosaic potexvirus satellite RNA is not essential for its replication and can be replaced with a bacterial gene.
A satellite RNA of 836 nt depends on the bamboo mosaic potexvirus (BaMV) for its replication and encapsulation. The BaMV satellite RNA (satBaMV) contains a single open reading frame encoding a 20-kDa nonstructural protein. A full-length infectious cDNA clone has been generated downstream of the T7 RNA polymerase promoter. To investigate the role of the 20-kDa protein encoded by satBaMV, satBaMV transcripts containing mutations in the open reading frame were tested for their ability to replicate in barley protoplasts and in Chenopodium quinoa using BaMV RNA as a helper genome. Unlike other large satellite RNAs, mutants in the open reading frame did not block their replication, suggesting that the 20-kDa protein is not essential for satBaMV replication. Precise replacement of the open reading frame with sequences encoding chloramphenicol acetyltransferase resulted in high level expression of chloramphenicol acetyltransferase in infected C. quinoa, indicating that satBaMV is potentially useful as a satellite-based expression vector.
Lin
N S
NS
Institute of Botany, Academia Sinica, Taipei, Taiwan, Republic of China.
Lee
Y S
YS
Lin
B Y
BY
Lee
C W
CW
Hsu
Y H
YH
D001483:Base Sequence; D002213:Capsid / Q000096:biosynthesis; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D017863:Potexvirus / Q000235:genetics / Q000502:physiology*; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012313:RNA / Q000096:biosynthesis / Q000235:genetics*; D019244:RNA, Satellite; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic; D014162:Transfection; D014758:Viral Core Proteins / Q000096:biosynthesis; D014764:Viral Proteins; D014779:Virus Replication
D017931:DNA Primers; D018076:DNA, Complementary; D019244:RNA, Satellite; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; D014764:Viral Proteins; D012313:RNA; D015500:Chloramphenicol O-Acetyltransferase; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases