Construction of a chimeric viral gene expressing plum pox virus coat protein.
The capsid-encoding gene of plum pox virus (PPV) was fused with the leader sequence of the coat protein mRNA (cp) of tobacco mosaic virus by a novel mutagenesis technique which involves reverse transcription of minus-strand RNA [synthesized by in vitro transcription of a double-stranded (ds) cDNA clone], using an ad hoc synthetic oligodeoxynucleotide as primer. The resulting cDNA was rendered ds and cloned into the plasmid, pBluescribe M13+. Transcription of this chimeric construction produced RNA molecules of 1250 nucleotides in length, which were used as messengers in the in vitro protein-synthesizing systems. The major product of this transcript consists of a 36-kDa polypeptide and was identified as the PPV coat protein (CP) by molecular weight estimation and by immunoprecipitation with a polyclonal antiserum to PPV. Transfer of this cDNA via Agrobacterium tumefaciens into plants was successfully performed. Transgenic Nicotiana plants producing the PPV CP were subsequently obtained.
Ravelonandro
M
M
Station de Pathologie Végétale, INRA, La Grande Ferrade, Villenave d'Ornon, France.
Monsion
M
M
Teycheney
P Y
PY
Delbos
R
R
Dunez
J
J
D001483:Base Sequence; D002213:Capsid / Q000235:genetics*; D002678:Chimera; D003001:Cloning, Molecular; D005814:Genes, Viral; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010942:Plant Viruses / Q000235:genetics*; D030821:Plants, Genetically Modified; D021382:Protein Sorting Signals / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015183:Restriction Mapping; D014027:Tobacco Mosaic Virus / Q000235:genetics; D014158:Transcription, Genetic
D021382:Protein Sorting Signals; D012333:RNA, Messenger