Selective recovery of foreign gene transcripts as virus-like particles in TMV-infected transgenic tobaccos.
A short origin-of-assembly sequence (OAS) located in the 30kDa movement protein gene, about 1.0kb from the 3'-end of the common strain of tobacco mosaic virus (TMV) RNA, nucleates encapsidation of the 6395-nucleotide-long genome by TMV coat protein in vitro, and presumably also in vivo. Single-stranded RNAs containing a foreign reporter gene sequence and the TMV OAS at their 5' - and 3' -ends, respectively, can be synthesized in vitro from recombinant SP6-transcription plasmids and will assemble spontaneously in vitro to form TMV-like 'pseudovirus' particles. In this paper, we show that foreign gene transcripts derived from the nuclear DNA of plants transformed by Agrobacterium tumefaciens, and which contain the TMV OAS, can be assembled into stable 'pseudovirus' particles in vivo during a systemic infection by TMV (helper). This is the first report of structural complementation between a heritable function bestowed on a transgenic plant and an infecting virus. As a route to protect, accumulate and recover a specific mRNA in vivo, in transgenic plant cells, this novel approach may find wider applications in developmental plant molecular biology.
D003001:Cloning, Molecular / Q000379:methods*; D005813:Genes, Synthetic; D005814:Genes, Viral; D005822:Genetic Vectors; D009024:Morphogenesis; D010947:Plants, Toxic; D012313:RNA / Q000235:genetics / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000302:isolation & purification; D012231:Rhizobium / Q000235:genetics; D014026:Nicotiana / Q000235:genetics*; D014027:Tobacco Mosaic Virus / Q000235:genetics* / Q000502:physiology; D014764:Viral Proteins / Q000502:physiology
D012333:RNA, Messenger; C039919:RNA, recombinant; D014764:Viral Proteins; D012313:RNA