Potato virus X as a vector for gene expression in plants.
The suitability of potato virus X (PVX) as a gene vector in plants was tested by analysis of two viral constructs. In the first, the GUS gene of Escherichia coli was substituted for the viral coat protein gene. In the second, GUS was added into the viral genome coupled to a duplicated copy of the viral promoter for the coat protein mRNA. The viral construct with the substituted coat protein gene accumulated poorly in inoculated protoplasts and failed to spread from the site of infection in plants. These results suggest a role for the viral coat protein in key stages of the viral infection cycle and show that gene replacement constructs are not suitable for the production of PVX-based gene vector. The construct with GUS coupled to the duplicated promoter for coat protein mRNA also accumulated less well in protoplasts than the unmodified PVX, but did infect systemically and directed high level synthesis of GUS in inoculated and systemically infected tissue. Although there was some genome instability in the PVX construct, much of the viral RNA in the systemically infected tissue had retained the foreign gene insertion, especially in infected Nicotiana clevelandii plants. These data point to a general utility of PVX as a vector for unregulated gene expression in plants.
Sainsbury Laboratory, Norwich Research Park, UK.
S
S
Chapman
T
T
Kavanagh
D
D
Baulcombe
D001483:Base Sequence; D015152:Blotting, Northern; D002213:Capsid / Q000235:genetics; D003062:Codon; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors; D016679:Genome, Viral; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010944:Plants / Q000235:genetics*; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D017863:Potexvirus / Q000235:genetics / Q000502:physiology; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000378:metabolism; D012367:RNA, Viral / Q000096:biosynthesis / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D003062:Codon; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D012367:RNA, Viral; D005966:Glucuronidase